bluuebunny/crossref_metadata_2025 huggingface dataset:
Looking at Patent Law: Patenting an Invention for Electroreduction of Carbon Dioxide and Electrooxidation of Hydrocarbons: A Case Study…Procedure for Continuing Prosecution of a Patent Application After Final Rejection…
Authors: Jennings Taylor E. , Inman Maria
Published: April 2024

In this installment of the “Looking at Patent Law” series, we present a case study of the prosecution events of U.S. Patent No. 11,788,193 directed towards simultaneous “Electroreduction of Carbon Dioxide and Electrooxidation of Hydrocarbons." The subject invention aligns with the research interest in sustainability matters by The Electrochemical Society and its members. The case study begins with a brief synopsis of the background of the invention followed by 1) summary of the drawing and the specification of the invention, 2) inventor assignment and power of attorney designations, 3) submission of the Invention Disclosure Statement (IDS) and associated Duty of Candor, 4) summary of the multiple non-final/final office actions rejecting the patent application, and 5) applicant response and allowance of the patent application. The case study illustrates the use of Request for Continued Examination (RCE) to continue the prosecution of the subject patent application after the USPTO issues a final rejection. With this case study, we hope to de-mystify the patent prosecution process and better prepare electrochemical and solid-state scientists, engineers and technologists to interact with their patent counsel regarding their inventions.
Effect of Organic Solvent on the Physical Properties of Spin-Coated Tin Sulfide Thin Films
Authors: Garmim T. , El Boughdadi M. , Benaissa N. , Soussi L. , Rhalmi O. , El Jouad Z. , Louardi A. , El Bachiri A. , Hartiti B. , Monkade M.
Published: April 2024

In the current study, the sol-gel spin coating was used to deposit SnS thin films on ordinary glass substrates, and annealed without sulfurization at 500 °C. Different organic solvents such as ethanol, 2-methoxyethanol, and methanol were used and their influence on the properties of SnS thin films was investigated. X-ray diffraction showed that the three prepared films have an orthorhombic structure with preferred orientation along (111) plan. In addition, the obtained films from ethanol solvent present better crystallinity with larger grain sizes around to 28.19 nm. Morphological and compositional analysis was obtained with a scanning electron microscope associated with an X-ray spectrometer. The results show better texture with uniform coverage on the glass substrate of the deposited films obtained for ethanol as solvent. The optical analysis obtained from the UV–vis spectrophotometer reveals that the deposited films show a broad absorption spectrum in the visible region and the band gap was found to lie in the range of 1.51 to 1.95 eV. The four-points probe measurement revealed smaller electrical resistivity for the films obtained from ethanol 1.76 × 102(Ω.cm) and a higher conductivity 5.68 × 10−3(Ω.cm)−1.
Structural and Electrical Properties of Li Substituted Mg Nano Ferrites: Prepared by Citrate-gel Auto Combustion Method
Authors: D. Mallesh , Naresh Pallati , Thalari Gangadhar , Seema Aravind
Published: April 2024

LixMg1−xFe2O4 where

x
=
0.0

to

0.5

with

0.1

nano ferrites were synthesized by citrate gel auto combustion. Using a field-emission scanning electron microscope (FE-SEM) and the X-ray diffraction technique, their structural and topographical studies were examined. The prepared nano ferrites’ single-phase cubic spinel along the Fd3m space group is confirmed by the structural investigation. The synthesized samples were found to have a crystalline size of 20–26 nm, as determined by the Debye–Scherrer formula. Lattice parameter variation observed for the samples indicates dopant atoms into a crystal lattice can also lead to changes in the lattice constant. FE-SEM images revealed the clear grain and grain boundaries with agglomerated structure and it varied between 33.6 nm-24.4 nm. Two bands observed in Fourier transform infrared spectroscopy illustrates the M-O band in tetrahedral and octahedral sites. Dielectric parameters of the sample observed by LCR meter. Single semicircle behavior observed in impedance spectra. Dielectric constant and dielectric loss decreased as increasing frequency explained by Koop’s theory of mechanism. AC conductivity increasing trend observed with frequency, mobility of charge carrier active at high frequency domain.
Aryl hydrocarbon receptor activity in intestinal epithelial cells in the formation of colonic tertiary lymphoid tissues
Authors: Garcia-Villatoro E. L. , Ufondu A. , Callaway E. S. , Allred K. F. , Safe S. H. , Chapkin R. S. , Jayaraman A. , Allred C. D.
Published: April 2024

This is the first research of its kind to demonstrate a clear connection between biological sex and the development of tertiary lymphoid tissues (TLT) in the colon. In addition, the research finds that in a preclinical model of inflammatory bowel disease, the expression of the aryl hydrocarbon receptor (AhR) influences the development of these structures in a sex-specific manner.
Condition Monitoring and Quantitative Evaluation of Railway Bridge Substructures Using Vehicle‐Induced Vibration Responses by Sparse Measurement
Authors: Wang Chuang , Zhan Jiawang , Wang Yujie , Xu Xinxiang , Wang Zhihang
Published: April 2024

Bridge substructure failure has been responsible for numerous recorded bridge collapses, particularly for small‐ and medium‐span bridges, so it is crucial to effectively monitor the performance of the bridge substructures for efficient maintenance and management. The current vibration‐based approaches for quantitatively evaluating bridge substructures rely on in‐situ experiments with a multitude of sensors or impact vibration test, making it challenging to implement long‐term online monitoring. This paper proposes an accurate, low cost, and practicable method to achieve online quantitative monitoring of railway bridge substructures using only one vibration sensor and operational train‐induced vibration responses. The newly derived flexible‐base Timoshenko beam models, along with the random decrement technique and Levenberg–Marquardt–Fletcher algorithm, are employed to identify the modal parameters and quantitatively assess the condition of bridge substructures. The proposed method is numerically verified through an established 3D train‐bridge‐foundation coupling system considering different damage scenarios. In addition, a real‐world application is also conducted on the 2nd Songhua River bridge in the Harbin–Dalian high‐speed railway, aiming at examining the effectiveness and robustness of the method in condition monitoring of bridge substructure under a complete freeze‐thaw cycle. The results indicate that the proposed methodology is effective in extracting the modal parameters and monitoring the state evolution of the bridge substructures, which offers an efficient and accurate strategy for condition monitoring and quantitative evaluation of railway bridge substructures.
Tarragon (Artemisia dracunculus) Essential Oil at Optimized Dietary Levels Prompted Growth, Immunity, and Resistance to Enteric Red‐Mouth Disease in the Rainbow Trout (Oncorhynchus mykiss)
Authors: Hajirezaee Saeed , Khanjani Mohammad Hossein , Ahani Saman , Ghiasvand Zahra
Published: April 2024

Fingerlings of the rainbow trout, Oncorhynchus mykiss (n = 300, 10.63 ± 0.6 g), were fed tarragon (Artemisia dracunculus) essential oil (TGO) for 2 months to examine its effects on growth properties, immunity, and resistance to Yersinia ruckeri infection. The treatments were control or TG1, TG2 (fed 0.5% TGO), TG3 (1% TGO), and TG4 (2% TGO). According to the results, an improvement was observed in growth parameters in all TGO‐treated groups compared to the control (P < 0.05). The digestive enzyme activities (protease and lipase) were significantly elevated in response to dietary TGO (P < 0.05). The immune system of the fish was enhanced by TGO, as it stimulated the immune parameters in serum (lysozyme, myeloperoxidase (MPO), alternative complement (ACH50), Ig) and mucus (lysozyme, protease, ACH50, Ig) (P < 0.05). The treatments, TG3 and TG4, showed more immune performance in response to TGO (P < 0.05). The fish in TG2 treatment had a higher levels of serum total protein than other groups (P < 0.05). The concentration of triglycerides (TRIG) and cholesterol (CHOL) in serum significantly decreased (P < 0.05) in response to TGO, as the lowest levels were observed in the treatment, TG3. The antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)) of serum elevated in TGO‐treated fish, with the maximum values for the TG4 group (P < 0.05). TGO reduced (P < 0.05) alanine aminotransferase (ALT) and alkaline phosphatase (ALP) levels in serum. After bacterial challenge, the TGO‐treated fish showed lower mortality compared to the control, where the lowest mortality was observed in TG4 (P < 0.05). In conclusion, TGO improved growth, immunity, and survival after bacterial challenge in the rainbow trout, with more performance in fish fed 1%–2% TGO.
Pore‐Scale Modeling of Pressure‐Driven Flow and Spontaneous Imbibition in Fracturing‐Shut‐In‐Flowback Process of Tight Oil Reservoirs
Authors: Jia Ninghong , Lv Weifeng , Liu Qingjie , Wang Daigang , Liu Fangzhou , Hu Zhe
Published: April 2024

Tight oil reservoirs are characterized by multiple pore spaces where nano‐micropores and multiscale fractures coexist, and each type of medium varies in scale, implying a tight coupling of multiscale fractures with matrix and giving rise to extremely complicated flow patterns. To further investigate its flow mechanism, we first construct three two‐dimensional (2‐D) fracture‐pore geometry models based on microfocus computed tomography (CT) imaging of a typical tight rock. A pore‐scale modeling workflow is thereafter developed using the Shan‐Chen lattice Boltzmann model (SC‐LBM) to simulate the pressure‐driven flow and spontaneous imbibition. The influence of fracture‐pore geometry on the pore‐scale fluid exchange dynamics in the fracturing‐shut‐in‐flowback process is clearly clarified. Results show that, for the porous medium model without fracture, the fracturing fluid can displace and replace some crude oil by spontaneous imbibition while a large amount of crude oil droplets remains unexploited away from the oil/water contact line, resulting in a low oil imbibition recovery. The injected fracturing fluid migrates into the deep position along the fractures, only a few entering the matrix pore space near the fractures. Small pores are the main channel for fracturing fluid to imbibe into the matrix pores, and the replaced crude oil droplets flow into fractures through large pores as intermittent or continuous pipe flow. The complexity of fracture network typically exerts a significant impact on the fluid exchange dynamics during pressure‐driven flow and spontaneous imbibition. The more complex the fracture network, the larger the volume of fracturing fluid injected, the easier for oil droplets replaced from the matrix pores, and the more difficult for the flowback of fracturing fluid in the fracture‐pore geometry model. Our understanding will provide a basis for explaining the underlying mechanisms of oil replacement by pressure‐driven flow and spontaneous imbibition in the fracturing‐shut‐in‐flowback process.
Study on Sound Absorption Properties of Polyvinyl Chloride (PVC) Film Multicavity Structure Materials
Authors: Zhang Tingying , Zhang Jiyang , Zheng Pengxuan , Hou Hong , Xu Ying
Published: April 2024

Since the development of industry, sound absorption and noise reduction have gradually become an urgent problem to be solved. Lightweight polymer film materials are very effective in response to sound waves, and sound waves can easily cause vibration of the film, which can convert sound energy into vibration and film friction to achieve sound absorption. The application conditions of the film material are very harsh, that is, a support body is required to fix the film and the film needs to be tensioned. The film is very thin and easy to damage. The idea of this research is to transform the film into a bubble structure and use a large number of film bubbles to form a cavity structure material. As a unit of the sound absorption structure, bubbles can avoid damage to the film. In this paper, commercial polyvinyl chloride film bubble materials are used to prepare two kinds of film multicavity structure materials, and the sound absorption performance of this film multicavity structure material is studied. The research results show that this film multicavity structure material has very excellent broadband sound absorption performance, which changes the narrow band sound absorption properties of the usual film single cavity. The average sound absorption coefficient can reach 0.84 in frequency range from 500 Hz to 6400 Hz. This structural material has a single peak sound absorption curve at the middle and low frequency bands, which is the characteristic of resonance sound absorption. And at the middle and high frequency bands, it exhibits the characteristics of broadband sound absorption. The film multicavity structure material has both cavity sound absorption and broadband sound absorption characteristics.
Heterogeneous Brain Atrophy Sites in Anxiety Disorders Map to a Common Brain Network
Authors: Yang Yinian , Xu Wenqiang , Wang Yingru , Cao Hai , Yao Xiaoqing , Zhang Ting , Xie Xiaohui , Hua Qiang , Cheng Wen , Shen Longshan , He Kongliang , Tian Yanghua , Wang Kai , Ji Gong-Jun
Published: April 2024

Background. Heterogeneous findings among anxiety disorder studies have hindered elucidation of the underlying pathophysiology and the development of mechanism-based therapies. Purpose. To determine whether structural MRI findings in anxiety disorder studies converge on a common network with therapeutic significance. Materials and Methods. In this retrospective study, a systematic literature search of PubMed and Web of Science databases was performed to identify coordinates of gray matter atrophy in patients with anxiety disorder. Atrophy coordinates were then mapped to an anxiety network constructed from the resting-state functional MRI (rs-fMRI) data of 652 healthy participants using “coordinate network mapping” and validated by specificity tests. The causal association of this network to anxiety symptoms was tested in a cohort of patients with brain lesions and emergent anxiety symptoms. The potential therapeutic utility of this anxiety network was then assessed by examining the clinical efficacy of network-targeted repetitive transcranial magnetic stimulation (rTMS) among a separate anxiety disorder cohort. Statistical analyses of images were performed using nonparametric tests and corrected for family-wise error. Results. Sixteen studies comprising 453 patients with anxiety (245 females; mean age±SD, 31.4±8.71 years) and 460 healthy controls (238 females; 31.7±10.08 years) were included in the analysis. Atrophy coordinates were mapped to an anxiety network with a hub region situated primarily within the superficial amygdala. Lesions associated with emergent anxiety symptoms exhibited stronger connectivity within this anxiety network than lesions not associated with anxiety (t=2.99; P=.004). Moreover, the connectivity strength of rTMS targets in the anxiety network was correlated with the improvements of anxiety symptom after treatment (r=.42, P=.02). Conclusions. Heterogeneous gray matter atrophy among patients with anxiety disorder localize to a common network that may serve as an effective therapeutic target.
Glucometabolic-Related Genes as Diagnostic Biomarkers and Therapeutic Targets for Alzheimer’s Disease and Type 2 Diabetes Mellitus: A Bioinformatics Analysis
Authors: Liu Shuo , Chen He , He Xiao-Dong , Yang Xiao-Ou
Published: April 2024

Background. Alzheimer’s disease (AD) and type 2 diabetes mellitus (T2DM) are two widespread chronic disorders characterized by shared risk factors and molecular pathways. Glucose metabolism, pivotal for cellular homeostasis and energy supply, plays a critical role in these diseases. Its disturbance has been linked to the pathogenesis of both AD and T2DM. However, a comprehensive investigation into the specific roles of glucometabolic genes in the onset and progression of AD and T2DM has yet to be conducted. Methods. By analyzing microarray datasets from the Gene Expression Omnibus (GEO) repository, we identified differentially expressed glucometabolic genes (DEGs) in AD and T2DM cohorts. A range of bioinformatics tools were employed for functional annotation, pathway enrichment, protein interaction network construction, module analysis, ROC curve assessment, correlation matrix construction, gene set enrichment analysis, and gene-drug interaction mapping of these DEGs. Key genes were further validated using quantitative real-time polymerase chain reaction (qRT-PCR) in AD and T2DM murine models. Results. Our investigation identified 41 glucometabolic-related DEGs, with six prominent genes (G6PD, PKM, ENO3, PFKL, PGD, and TALDO1) being common in both AD and T2DM cohorts. These genes play crucial roles in metabolic pathways including glycolysis, pentose phosphate pathway, and amino sugar metabolism. Their diagnostic potential was highlighted by area under curve (AUC) values exceeding 0.6 for AD and 0.8 for T2DM. Further analysis explored the interactions, pathway enrichments, regulatory mechanisms, and potential drug interactions of these key genes. In the AD murine model, quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed significant upregulation of G6pd, Eno3, and Taldo1. Similarly, in the T2DM murine model, elevated expression levels of G6pd, Pfkl, Eno3, and Pgd were observed. Conclusion. Our rigorous research sheds light on the molecular interconnections between AD and T2DM from a glucometabolic perspective, revealing new opportunities for pharmacological innovation and therapeutic approaches. This study appears to be the first to extensively investigate glucometabolic-associated DEGs and key genes in both AD and T2DM, utilizing multiple datasets. These insights are set to enhance our understanding of the complex pathophysiology underlying these widespread chronic diseases.
TSH Receptor Reduces Hemoglobin S Polymerization and Increases Deformability and Adhesion of Sickle Erythrocytes
Authors: Mendonça-Reis Evelyn , Guimarães-Nobre Camila Cristina , Teixeira-Alves Lyzes Rosa , Miranda-Alves Leandro , Berto-Junior Clemilson
Published: April 2024

SCD is a hereditary disorder caused by genetic mutation in the beta-globin gene, resulting in abnormal hemoglobin, HbS that forms sickle-shaped erythrocytes under hypoxia. Patients with SCD have endocrine disorders and it was described that 7% of these patients have clinical hypothyroidism. Recent studies have shown that mature erythrocytes possess TSH receptors. Thus, we aimed to assess the effects of TSH on SCD erythrocytes. The experiments were conducted using different concentrations of TSH (1, 2, 3, and 5 mIU/L). In HbS polymerization assay, erythrocytes were exposed to TSH in hypoxia to induce polymerization, and measurements were taken for 30 minutes. The deformability assay was made using Sephacryl-S 500 columns to separate deformable from nondeformable cells. Static adhesion test utilized thrombospondin to assess erythrocyte adhesion in the presence of TSH. TSH at all contractions were able to reduce polymerization of HbS and increase deformability. The static adhesion of erythrocytes at the lowest concentrations of 1 and 2 mIU/L were increased, but at higher contractions of 3 and 5 mIU/L, static adhesion was not modulated. The results suggest that TSH has potential involvement in the pathophysiology of sickle cell disease by inhibiting HbS polymerization, positively modulating deformability and impacting static adhesion to thrombospondin.
Emotion Regulation as a Mechanism of Mindfulness in Individual Cognitive-Behavioral Therapy for Depression and Anxiety Disorders
Authors: Pruessner Luise , Timm Christina , Kalmar Julia , Bents Hinrich , Barnow Sven , Mander Johannes
Published: April 2024

Background. The global prevalence of depression and anxiety disorders underscores the need for a more profound comprehension of effective treatments. Mindfulness has shown promise in enhancing treatment outcomes and preventing relapse in these conditions, but the underlying mechanisms remain poorly understood. Methods. This study examined the role of emotion regulation as a mediator in the relationship between changes in mindfulness and the reduction of depression and anxiety symptoms during individual cognitive-behavioral therapy (CBT). We tracked longitudinal changes in mindfulness, emotion regulation, depression, and anxiety at baseline (pre), early treatment (session 5), midtreatment (session 15), and posttreatment (session 25) in 162 patients with depression and anxiety disorders. Further, we examined whether the effects of mindfulness on emotion regulation could be enhanced by introducing a brief mindfulness intervention at the beginning of each CBT session, as compared to progressive muscle relaxation and individual psychotherapy without any standardized session-introducing interventions. Results. Multilevel structural equation modeling indicated that decreases in rumination and increases in reappraisal and acceptance mediated the relationship between mindfulness and reductions in depressive symptoms. In contrast, reductions in avoidance explained the association between mindfulness and changes in anxiety symptoms. These links remained unchanged when adding a mindfulness intervention into individual CBT. Conclusion. Results support emotion regulatory properties of mindfulness and highlight distinct pathways of symptom reduction in depression and anxiety. These findings have important implications for understanding the mechanisms of mindfulness and tailoring treatment to individual patient needs. This trial is registered with NTC02270073.
A Case of Diltiazem-Induced Pustular Rash in an 83-Year-Old Female
Authors: Bangolo Ayrton I. , Yelton-Torres Karen , Mahajan Atharv , Patel Vrusha , Sajjad Laiba , Ansong Prince Ofori , Kashyap Monisha , Batura Umesh , Ravulapalli Madhavi , Perera Ayodya , Weissman Simcha
Published: April 2024

Acute generalized exanthematous pustulosis (AGEP) is a rare, acute skin eruption characterized by the development of numerous nonfollicular sterile pustules. Most cases are caused by drug reactions, among which Diltiazem has been incriminated. Herein, we present an 83-year-old female who presented for evaluation of generalized skin rash 3 days after initiation of Diltiazem. She was eventually diagnosed with AGEP, Diltiazem was discontinued, and systemic steroids were administered with the resolution of symptoms. This case report has the objective of encouraging clinicians to include AGEP in the differential diagnosis of skin eruption following the initiation of Diltiazem.
Supplementary Figure 8 from Elimusertib has Antitumor Activity in Preclinical Patient-Derived Pediatric Solid Tumor Models
Authors: Pusch Fabian F. , Dorado García Heathcliff , Xu Robin , Gürgen Dennis , Bei Yi , Brückner Lotte , Röefzaad Claudia , von Stebut Jennifer , Bardinet Victor , Chamorro Gonzalez Rocío , Eggert Angelika , Schulte Johannes H. , Hundsdörfer Patrick , Seifert Georg , Haase Kerstin , Schäfer Beat W. , Wachtel Marco , Kühl Anja A. , Ortiz Michael V. , Wengner Antje M. , Scheer Monika , Henssen Anton G.
Published: April 2024

Supplementary Figure 8 shows the IHC markers tested in EWS PDXs.
Supplementary Figure S9 from Combination Therapy with EGFR Tyrosine Kinase Inhibitors and TEAD Inhibitor Increases Tumor Suppression Effects in EGFR Mutation–positive Lung Cancer
Authors: Ogimoto Tatsuya , Ozasa Hiroaki , Tsuji Takahiro , Funazo Tomoko , Yamazoe Masatoshi , Hashimoto Kentaro , Yoshida Hiroshi , Hosoya Kazutaka , Ajimizu Hitomi , Nomizo Takashi , Yoshida Hironori , Hamaji Masatsugu , Menju Toshi , Yoshizawa Akihiko , Date Hiroshi , Hirai Toyohiro
Published: April 2024

Supplementary Figure S9 shows cell viability assays of PC9, HCC827, and H1975 cells treated with VT104.
Supplementary Figure S6 from Combination Therapy with EGFR Tyrosine Kinase Inhibitors and TEAD Inhibitor Increases Tumor Suppression Effects in EGFR Mutation–positive Lung Cancer
Authors: Ogimoto Tatsuya , Ozasa Hiroaki , Tsuji Takahiro , Funazo Tomoko , Yamazoe Masatoshi , Hashimoto Kentaro , Yoshida Hiroshi , Hosoya Kazutaka , Ajimizu Hitomi , Nomizo Takashi , Yoshida Hironori , Hamaji Masatsugu , Menju Toshi , Yoshizawa Akihiko , Date Hiroshi , Hirai Toyohiro
Published: April 2024

Supplementary Figure S6 shows the relative expression of CTGF in KTOR27 cells treated with afatinib with YAP1 knockdown.
Figure S7 from ADCT-602, a Novel PBD Dimer–containing Antibody–Drug Conjugate for Treating CD22-positive Hematologic Malignancies
Authors: Zammarchi Francesca , Havenith Karin E. , Sachini Nikoleta , Janghra Narinder , Chivers Simon , Idusogie Esohe , Gaudio Eugenio , Tarantelli Chiara , Bertelli Francois , Santos Kathleen , Tyrer Peter , Corbett Simon , Spriano Filippo , Golino Gaetanina , Cascione Luciano , Bertoni Francesco , Hartley John A. , van Berkel Patrick H.
Published: April 2024

Supplementary Fig. S7. Cynomolgus monkey body weight after dosing ADCT-602 at 0.6 or 0.9 mg/kg (2 doses, 21 days apart). Data presented as mean body weight (kg) ± SEM (each group, n = 3). SEM standard error of mean.
Supplementary Figure 3 from Oncogenic Cells of Renal Embryonic Lineage Sensitive to the Small-Molecule Inhibitor QC6352 Display Depletion of KDM4 Levels and Disruption of Ribosome Biogenesis
Authors: Pichavaram Prahalathan , Jablonowski Carolyn M. , Fang Jie , Fleming Andrew M. , Gil Hyea Jin , Boghossian Andrew S. , Rees Matthew G. , Ronan Melissa M. , Roth Jennifer A. , Morton Christopher L. , Zambetti Gerard P. , Davidoff Andrew M. , Yang Jun , Murphy Andrew J.
Published: April 2024

QC6352 does not cause a biologically significant increase in cell death or apoptosis in WiT49 or HEK293 cells. The percent difference in cell death was statistically significant in HEK293 cells, but the absolute difference was less than 1%. ***paired two-tailed t-test p value <0.001; ns=non-significant.
Supplementary Figure 14 from Oncogenic Cells of Renal Embryonic Lineage Sensitive to the Small-Molecule Inhibitor QC6352 Display Depletion of KDM4 Levels and Disruption of Ribosome Biogenesis
Authors: Pichavaram Prahalathan , Jablonowski Carolyn M. , Fang Jie , Fleming Andrew M. , Gil Hyea Jin , Boghossian Andrew S. , Rees Matthew G. , Ronan Melissa M. , Roth Jennifer A. , Morton Christopher L. , Zambetti Gerard P. , Davidoff Andrew M. , Yang Jun , Murphy Andrew J.
Published: April 2024

siRNA mediated knockdown of KDM4A, B, C, and KDM4A-C did not reduce ribosomal protein expression in WiT49 and HEK293 cells.
Supplementary Table S1 from Cereblon-based Bifunctional Degrader of SOS1, BTX-6654, Targets Multiple KRAS Mutations and Inhibits Tumor Growth
Authors: Begovich Kyle , Schoolmeesters Angela , Rajapakse Navin , Martinez-Terroba Elena , Kumar Maneesh , Shakya Arvind , Lai Chon , Greene Steven , Whitefield Brandon , Okano Akinori , Mali Venkat , Huang Shenlin , Chourasia Aparajita H. , Fung Leah
Published: April 2024

Supplementary Table 1 shows all the cell lines used in this study along with purchasing information as well as conditions for cell culture
Supplementary Figure S5 from Cereblon-based Bifunctional Degrader of SOS1, BTX-6654, Targets Multiple KRAS Mutations and Inhibits Tumor Growth
Authors: Begovich Kyle , Schoolmeesters Angela , Rajapakse Navin , Martinez-Terroba Elena , Kumar Maneesh , Shakya Arvind , Lai Chon , Greene Steven , Whitefield Brandon , Okano Akinori , Mali Venkat , Huang Shenlin , Chourasia Aparajita H. , Fung Leah
Published: April 2024

Supplementary Figure S5 demonstrates the synergistic effects of combining BTX-7312 with inhibitors of the RTK-RAS-MAPK pathway
Figure S10 from Neutrophil Elastase Remodels Mammary Tumors to Facilitate Lung Metastasis
Authors: Lulla Amriti R. , Akli Said , Karakas Cansu , Caruso Joseph A. , Warma Lucas D. , Fowlkes Natalie W. , Rao Xiayu , Wang Jing , Hunt Kelly K. , Watowich Stephanie S. , Keyomarsi Khandan
Published: April 2024

Supplementary Figure S10: Gating strategy used to identify myeloid, lymphoid and B-cell populations for the experiment described in Supplementary Figure S2. All analyses were conducted on FlowJo and statistical analyses were done on GraphPad Prism.
Table S2 from Development of a Novel DNA Mono-alkylator Platform for Antibody–Drug Conjugates
Authors: Thomas Joshua D. , Yurkovetskiy Aleksandr V. , Yin Mao , Bodyak Natalya D. , Tang Shuyi , Protopopova Marina , Kelleher Eugene , Jones Brian , Yang Liping , Custar Daniel , Catcott Kalli C. , Demady Damon R. , Collins Scott D. , Xu Ling , Bu Charlie , Qin LiuLiang , Ter-Ovanesyan Elena , Damelin Marc , Toader Dorin , Lowinger Timothy B.
Published: April 2024

MRM Quantification Parameters
Figure S5 from Development of a Novel DNA Mono-alkylator Platform for Antibody–Drug Conjugates
Authors: Thomas Joshua D. , Yurkovetskiy Aleksandr V. , Yin Mao , Bodyak Natalya D. , Tang Shuyi , Protopopova Marina , Kelleher Eugene , Jones Brian , Yang Liping , Custar Daniel , Catcott Kalli C. , Demady Damon R. , Collins Scott D. , Xu Ling , Bu Charlie , Qin LiuLiang , Ter-Ovanesyan Elena , Damelin Marc , Toader Dorin , Lowinger Timothy B.
Published: April 2024

Figure shows Plasma stability comparing indole-linked ADC T-13 to N10-linked ADC T-BH
Figure S3 from The Methyltransferases METTL7A and METTL7B Confer Resistance to Thiol-Based Histone Deacetylase Inhibitors
Authors: Robey Robert W. , Fitzsimmons Christina M. , Guiblet Wilfried M. , Frye William J.E. , González Dalmasy José M. , Wang Li , Russell Drake A. , Huff Lyn M. , Perciaccante Andrew J. , Ali-Rahmani Fatima , Lipsey Crystal C. , Wade Heidi M. , Mitchell Allison V. , Maligireddy Siddhardha S. , Terrero David , Butcher Donna , Edmondson Elijah F. , Jenkins Lisa M. , Nikitina Tatiana , Zhurkin Victor B. , Tiwari Amit K. , Piscopio Anthony D. , Totah Rheem A. , Bates Susan E. , Arda H. Efsun , Gottesman Michael M. , Batista Pedro J.
Published: April 2024

Figure S3 contains gene ontology analysis, motif analysis, and nucleosome density from the RNA-seq and ATAC-seq data.
Figure S4 from Mass Spectrometry–Based Proteomics Identifies Serpin B9 as a Key Protein in Promoting Bone Metastases in Lung Cancer
Authors: Huang Yufeng , Gong Ming , Chen Hongmin , Deng Chuangzhong , Zhu Xiaojun , Lin Jiaming , Huang Anfei , Xu Yanyang , Tai Yi , Song Guohui , Xu Huaiyuan , Hu Jinxin , Feng Huixiong , Tang Qinglian , Lu Jinchang , Wang Jin
Published: April 2024

Loss of SB9 can restore infiltration of CTLs.
A-B: Quantification of tumour volume size by BLI curve in indicated groups.Collected tumor tissues one week after occurrence of BM and selected the certain volume of tumor tissues as same as possible.
C: Flow cytometry analyses demonstrate that the infiltration of CD4+CD25+Foxp3+Tregs were no difference in BM from the SB9-silenced-group and SB9-overexpression-group.
D-E: Flow cytometry analyses demonstrated that the infiltration of CD44highCD62LlowCD8+ effector memory T cells(D), and TNF-α+CD8+ T cells (E) were significantly higher in BM from the LLC1-Pa-sr-ctrl and LLC1-BM3-shSB9 group than the LLC1-Pa-OE-SB9 and LLC1-BM3-sr-ctrl group, respectively.
F-G: Flow cytometry analyses show significantly lower percentages of pro-tumor-infiltrating IL-10 CD4+T cells(F) in LLC1-Pa-sr-ctrl and LLC1-BM3-shSB9 group than the LLC1-Pa-OE-SB9 and LLC1-BM3-sc-ctrl group, respectively. Moreover,anti-tumor-infiltrating IL-2 CD4+T(G) cells were significantly higher percentages in LLC1-Pa-sr-ctrl and LLC1-BM3-shSB9 group than the LLC1-Pa-OE-SB9 and LLC1-BM3-sr-ctrl group, respectively.
H: Flow cytometry analyses demonstrated that the exhausted state(LAG-3) infiltration of CD8+T cells were no difference in BM from the SB9-silenced-group and SB9-overexpression-group. *P<0.05; **P<0.01; ***P<0.001; ns: not significant; P values were obtained by two-tailed unpaired t-test; The data are displayed as an average ± s.e.m.
Figure S3 from Mass Spectrometry–Based Proteomics Identifies Serpin B9 as a Key Protein in Promoting Bone Metastases in Lung Cancer
Authors: Huang Yufeng , Gong Ming , Chen Hongmin , Deng Chuangzhong , Zhu Xiaojun , Lin Jiaming , Huang Anfei , Xu Yanyang , Tai Yi , Song Guohui , Xu Huaiyuan , Hu Jinxin , Feng Huixiong , Tang Qinglian , Lu Jinchang , Wang Jin
Published: April 2024

SB9 is a key protein in promoting BM and bone-related adverse events (SRE) in LC
A: Fracture frequency of the indicated mice (n = 10/group) from indicated experimental group.
B: The incidence of lung, brain and other organ metastases in indicated groups from immunocompetent C57BL/6 mice with IC injection. *P<0.05; **P<0.01; ***P<0.001; ns: not significant; P values were obtained by Two-sides with chi-square (and Fisher's exact) tests.
C-D: Cell clone formation assay. There was no difference in cell clone formation between LLC1-BM3-sr-ctrl vs LLC1-BM3-shSB9 and LLC1-Pa-sr-ctrl vs LLC1-Pa-OE-SB9 groups; hole plate photo(B); number of clone formation(B) (sr-ctrl：scramble control; OE:overexpression).
E: Ki67 mRNA levels in LLC1 derivative cell lines.There was no difference in Ki67 mRNA levels between LLC1-BM3-sr-ctrl vs LLC1-BM3-shSB9 and LLC1-Pa-sr-ctrl vs LLC1-Pa-OE-SB9 groups.
F: Flow cytometry analysis the proportion of mcherry positive cells in LLC1-BM3-OVA and LLC1-BM3-shSB9-OVA cells.
G: The incidence of lung, brain and other organ metastases in indicated groups from immunodeficient BABL/C nude mice with CA injection. *P<0.05; **P<0.01; ***P<0.001; ns: not significant; P values were obtained by Two-sides with chi-square (and Fisher's exact) tests.
Figure S4 from miR-217 Regulates Normal and Tumor Cell Fate Following Induction of Endoplasmic Reticulum Stress
Authors: Dey Neekkan , Koumenis Costas , Ruggero Davide , Fuchs Serge Y. , Diehl J. Alan
Published: April 2024

S4. (a) The 3’UTR of SUZ12 mRNA contains the binding sequence of miR-217 as predicted by TargetScan. (b) Map of luciferase reporter vector Wt-SUZ12-3’UTR (HmiT088547-MT06, GeneCopoeia).
Figure S2 from NRAS Mutant Dictates AHCYL1-Governed ER Calcium Homeostasis for Melanoma Tumor Growth
Authors: Cai Chufan , Tu Jiayi , Najarro Jeronimo , Zhang Rukang , Fan Hao , Zhang Freya Q. , Li Jiacheng , Xie Zhicheng , Su Rui , Dong Lei , Arellano Nicole , Ciboddo Michele , Elf Shannon E. , Gao Xue , Chen Jing , Wu Rong
Published: April 2024

Supplementary Figure 2. AHCYL1 knockout selectively causes cell cycle arrest in NRAS-mutant expressing human melanoma cells.
IL2 Targeted to CD8+ T Cells Promotes Robust Effector T-cell Responses and Potent Antitumor Immunity
Authors: Moynihan Kelly D. , Kumar Manu P. , Sultan Hussein , Pappas Danielle C. , Park Terrence , Chin S. Michael , Bessette Paul , Lan Ruth Y. , Nguyen Henry C. , Mathewson Nathan D. , Ni Irene , Chen Wei , Lee Yonghee , Liao-Chan Sindy , Chen Jessie , Schumacher Ton N.M. , Schreiber Robert D. , Yeung Yik A. , Djuretic Ivana M.
Published: April 2024

IL2 signals pleiotropically on diverse cell types, some of which contribute to therapeutic activity against tumors, whereas others drive undesired activity, such as immunosuppression or toxicity. We explored the theory that targeting of IL2 to CD8+ T cells, which are key antitumor effectors, could enhance its therapeutic index. To this aim, we developed AB248, a CD8 cis-targeted IL2 that demonstrates over 500-fold preference for CD8+ T cells over natural killer and regulatory T cells (Tregs), which may contribute to toxicity and immunosuppression, respectively. AB248 recapitulated IL2’s effects on CD8+ T cells in vitro and induced selective expansion of CD8+T cells in primates. In mice, an AB248 surrogate demonstrated superior antitumor activity and enhanced tolerability as compared with an untargeted IL2Rβγ agonist. Efficacy was associated with the expansion and phenotypic enhancement of tumor-infiltrating CD8+ T cells, including the emergence of a “better effector” population. These data support the potential utility of AB248 in clinical settings.

Significance: The full potential of IL2 therapy remains to be unlocked. We demonstrate that toxicity can be decoupled from antitumor activity in preclinical models by limiting IL2 signaling to CD8+ T cells, supporting the development of CD8+ T cell–selective IL2 for the treatment of cancer.

See related article by Kaptein et al. p. 1226.
Supplementary Fig. S7 from Reprogramming the Intrahepatic Cholangiocarcinoma Immune Microenvironment by Chemotherapy and CTLA-4 Blockade Enhances Anti–PD-1 Therapy
Authors: Chen Jiang , Amoozgar Zohreh , Liu Xin , Aoki Shuichi , Liu Zelong , Shin Sarah M. , Matsui Aya , Hernandez Alexei , Pu Zhangya , Halvorsen Stefan , Lei Pin-Ji , Datta Meenal , Zhu Lingling , Ruan Zhiping , Shi Lei , Staiculescu Daniel , Inoue Koetsu , Munn Lance L. , Fukumura Dai , Huang Peigen , Sassi Slim , Bardeesy Nabeel , Ho Won Jin , Jain Rakesh K. , Duda Dan G.
Published: April 2024

Supplementary Fig. S7: Immunophenotyping in murine 425-ICC and SS49-ICC.
Supplementary Figure 7 from The Immune Suppressor IGSF1 as a Potential Target for Cancer Immunotherapy
Authors: Koh Dong-In , Lee Minki , Park Yoon Sun , Shin Jae-Sik , Kim Joseph , Ryu Yea Seong , Lee Jun Hyung , Bae Seunggeon , Lee Mi So , Hong Jun Ki , Jeong Hong-Rae , Choi Mingee , Hong Seung-Woo , Kim Dong Kwan , Lee Hyun-kyung , Kim Bomi , Yoon Yoo Sang , Jin Dong-Hoon
Published: April 2024

WM-A1 induces T cell activation in vitro and in vivo.
Supplementary Figure 5 from The Immune Suppressor IGSF1 as a Potential Target for Cancer Immunotherapy
Authors: Koh Dong-In , Lee Minki , Park Yoon Sun , Shin Jae-Sik , Kim Joseph , Ryu Yea Seong , Lee Jun Hyung , Bae Seunggeon , Lee Mi So , Hong Jun Ki , Jeong Hong-Rae , Choi Mingee , Hong Seung-Woo , Kim Dong Kwan , Lee Hyun-kyung , Kim Bomi , Yoon Yoo Sang , Jin Dong-Hoon
Published: April 2024

Target identification was conducted to identify the counter-partner of IGSF1.
Supplementary Figure 5 from Tumor Burden Dictates the Neoantigen Features Required to Generate an Effective Cancer Vaccine
Authors: Garzia Irene , Nocchi Linda , Avalle Lidia , Troise Fulvia , Leoni Guido , Seclì Laura , Antonucci Laura , Cotugno Gabriella , Allocca Simona , Romano Giuseppina , Conti Laura , Caiazza Carmen , Mallardo Massimo , Poli Valeria , Scarselli Elisa , D'Alise Anna Morena
Published: April 2024

Representative numbers of total and #23+ CD8+ T cells in tumor and spleen of Help versus non Help vaccine. A) Experimental scheme, FACS gating strategy with representative FACS plots of intratumoral #23+ CD8+ T cells in Help group and untreated group. B and C) Average number of total CD8+ and #23+ T cells relative to figure 6B, 6C, 6E, 6F, per each experimental group (Untreated, Help and No Help) in tumor (B) and in spleen (C) at day 14 post immunization. Number (n) of total CD8+ (x104 cells) and Ag-#23+ specific (x102 cells). Data are shown as total cell count per tumor / organ.
Supplementary Figure 2 from Immune Modulation with RANKL Blockade through Denosumab Treatment in Patients with Cancer
Authors: Chang Hewitt , Marquez Jaqueline , Chen Brandon K. , Kim Daniel M. , Cheng Michael L. , Liu Eric V. , Yang Hai , Zhang Li , Sinha Meenal , Cheung Alexander , Kwek Serena S. , Chow Eric D. , Bridge Mark , Aggarwal Rahul R. , Friedlander Terence W. , Small Eric J. , Anderson Mark , Fong Lawrence
Published: April 2024

Supplementary Figure 2: TREC and RANKL quantitation
(A) Standard curves of tRNA were prepared on every reaction plate in triplicate for 100,000 to 12 TREC copies and -actin. (B) Gene expression of RANKL in circulating immune cells was quantified by qPCR. Fold-change expression of RANKL is relative to TP1. (C) Concentration of soluble RANKL from plasma was quantified at pretreatment TP1 and at TP3. (D) Gene expression of RANKL was compared between concurrent therapy groups. Fold-change expression of RANKL is relative to TP1.
Supplementary Table 1 from Gut Mycobiota Dysbiosis Is Associated with Melanoma and Response to Anti–PD-1 Therapy
Authors: Szóstak Natalia , Handschuh Luiza , Samelak-Czajka Anna , Tomela Katarzyna , Pietrzak Bernadeta , Schmidt Marcin , Galus Łukasz , Mackiewicz Jacek , Mackiewicz Andrzej , Kozlowski Piotr , Philips Anna
Published: April 2024

Supplementary Table 1. Melanoma patients and healthy controls data.
Supplementary Figure 4 from Gut Mycobiota Dysbiosis Is Associated with Melanoma and Response to Anti–PD-1 Therapy
Authors: Szóstak Natalia , Handschuh Luiza , Samelak-Czajka Anna , Tomela Katarzyna , Pietrzak Bernadeta , Schmidt Marcin , Galus Łukasz , Mackiewicz Jacek , Mackiewicz Andrzej , Kozlowski Piotr , Philips Anna
Published: April 2024

Supplementary Figure 4. Phylogenetic trees of mycobiota in (A) healthy and (B) melanoma subjects.
Supplementary Table 1 from Neutrophils Mediate Protection Against Colitis and Carcinogenesis by Controlling Bacterial Invasion and IL22 Production by γδ T Cells
Authors: Carnevale Silvia , Ponzetta Andrea , Rigatelli Anna , Carriero Roberta , Puccio Simone , Supino Domenico , Grieco Giovanna , Molisso Piera , Di Ceglie Irene , Scavello Francesco , Perucchini Chiara , Pasqualini Fabio , Recordati Camilla , Tripodo Claudio , Belmonte Beatrice , Mariancini Andrea , Kunderfranco Paolo , Sciumè Giuseppe , Lugli Enrico , Bonavita Eduardo , Magrini Elena , Garlanda Cecilia , Mantovani Alberto , Jaillon Sebastien
Published: April 2024

Supplementary Table 1. Colitis grade score.
Supplementary Fig. S1 from Reprogramming the Intrahepatic Cholangiocarcinoma Immune Microenvironment by Chemotherapy and CTLA-4 Blockade Enhances Anti–PD-1 Therapy
Authors: Chen Jiang , Amoozgar Zohreh , Liu Xin , Aoki Shuichi , Liu Zelong , Shin Sarah M. , Matsui Aya , Hernandez Alexei , Pu Zhangya , Halvorsen Stefan , Lei Pin-Ji , Datta Meenal , Zhu Lingling , Ruan Zhiping , Shi Lei , Staiculescu Daniel , Inoue Koetsu , Munn Lance L. , Fukumura Dai , Huang Peigen , Sassi Slim , Bardeesy Nabeel , Ho Won Jin , Jain Rakesh K. , Duda Dan G.
Published: April 2024

Supplementary Fig. S1: Gating strategies for flow cytometry.
Supplementary Data from Reprogramming the Intrahepatic Cholangiocarcinoma Immune Microenvironment by Chemotherapy and CTLA-4 Blockade Enhances Anti–PD-1 Therapy
Authors: Chen Jiang , Amoozgar Zohreh , Liu Xin , Aoki Shuichi , Liu Zelong , Shin Sarah M. , Matsui Aya , Hernandez Alexei , Pu Zhangya , Halvorsen Stefan , Lei Pin-Ji , Datta Meenal , Zhu Lingling , Ruan Zhiping , Shi Lei , Staiculescu Daniel , Inoue Koetsu , Munn Lance L. , Fukumura Dai , Huang Peigen , Sassi Slim , Bardeesy Nabeel , Ho Won Jin , Jain Rakesh K. , Duda Dan G.
Published: April 2024

RNAseq dataset
Data from Tumor Burden Dictates the Neoantigen Features Required to Generate an Effective Cancer Vaccine
Authors: Garzia Irene , Nocchi Linda , Avalle Lidia , Troise Fulvia , Leoni Guido , Seclì Laura , Antonucci Laura , Cotugno Gabriella , Allocca Simona , Romano Giuseppina , Conti Laura , Caiazza Carmen , Mallardo Massimo , Poli Valeria , Scarselli Elisa , D'Alise Anna Morena
Published: April 2024

Abstract
Tumor neoantigens (nAg) represent a promising target for cancer immunotherapy. The identification of nAgs that can generate T-cell responses and have therapeutic activity has been challenging. Here, we sought to unravel the features of nAgs required to induce tumor rejection. We selected clinically validated Great Ape–derived adenoviral vectors (GAd) as a nAg delivery system for differing numbers and combinations of nAgs. We assessed their immunogenicity and efficacy in murine models of low to high disease burden, comparing multi-epitope versus mono-epitope vaccines. We demonstrated that the breadth of immune response is critical for vaccine efficacy and having multiple immunogenic nAgs encoded in a single vaccine improves efficacy. The contribution of each single neoantigen was examined, leading to the identification of 2 nAgs able to induce CD8+ T cell–mediated tumor rejection. They were both active as individual nAgs in a setting of prophylactic vaccination, although to different extents. However, the efficacy of these single nAgs was lost in a setting of therapeutic vaccination in tumor-bearing mice. The presence of CD4+ T-cell help restored the efficacy for only the most expressed of the two nAgs, demonstrating a key role for CD4+ T cells in sustaining CD8+ T-cell responses and the necessity of an efficient recognition of the targeted epitopes on cancer cells by CD8+ T cells for an effective antitumor response. This study provides insight into understanding the determinants of nAgs relevant for effective treatment and highlights features that could contribute to more effective antitumor vaccines.

See related Spotlight by Slingluff Jr, p. 382.
CD4+ Regulatory T Cells in Human Cancer: Subsets, Origin, and Molecular Regulation
Authors: Swatler Julian , De Luca Marco , Rotella Ivano , Lise Veronica , Mazza Emilia Maria Cristina , Lugli Enrico
Published: April 2024

CD4+CD25hiFOXP3+ regulatory T cells (Treg) play major roles in the maintenance of immune tolerance, prevention of inflammation, and tissue homeostasis and repair. In contrast with these beneficial roles, Tregs are abundant in virtually all tumors and have been mechanistically linked to disease progression, metastases development, and therapy resistance. Tregs are thus recognized as a major target for cancer immunotherapy. Compared with other sites in the body, tumors harbor hyperactivated Treg subsets whose molecular characteristics are only beginning to be elucidated. Here, we describe current knowledge of intratumoral Tregs and discuss their potential cellular and tissue origin. Furthermore, we describe currently recognized molecular regulators that drive differentiation and maintenance of Tregs in cancer, with a special focus on those signals regulating their chronic immune activation, with relevant implications for cancer progression and therapy.
Optimal Neoantigen Cancer Vaccines Target CD8+ and CD4+ T Cells with Multiple Antigens
Authors: Slingluff Craig L.
Published: April 2024

Cancer vaccines targeting mutated neoantigens offer promise for prevention of cancer recurrence and for treatment of established cancers. Questions remain about whether vaccines need to target multiple neoantigens and whether they need to target both CD8+ and CD4+ T cells. In this issue, Garzia and colleagues demonstrate the importance of including multiple antigens to stimulate both CD4+ T cells and CD8+ T cells for treatment of established cancer.

See related article by Garzia et al., p. 440 (4).
Allogeneic CAR T Cells: Complex Cellular Therapy Designs Test the Limits of Our Preclinical Models
Authors: Caimi Paolo F. , Melenhorst Jan Joseph
Published: April 2024

All chimeric antigen receptor (CAR) T-cell products currently approved by the FDA are autologous, which poses several challenges for widespread use. In this issue, Degagné and colleagues present their preclinical research on creating off-the-shelf CAR T cells for multiple myeloma. They utilized the CRISPR/Cas12a genome editing platform and gene knock-in techniques to eliminate alloreactivity and decrease susceptibility to natural killer (NK)-cell elimination. This work has led to an ongoing phase I trial of off-the-shelf CAR T cells for multiple myeloma treatment.

See related article by Degagné et al., p. 462 (2).
FIGURE 8 from Identification of Enhanced Vaccine Mimotopes for the p15E Murine Cancer Antigen
Authors: Zhou Shiqi , Song Yiting , Luo Yuan , Quinn Breandan , Jiao Yang , Long Mark D. , Abrams Scott I. , Lovell Jonathan F.
Published: April 2024

TCR sequencing of wild-type p15E-specific CD8 T cells sorted from splenocytes of 3C, 3M, and 3C2V immunized mice. A, Percentage of Ag-specific CD8+ T cells in all CD8+ T cells. B, Proportion of total repertoire occupied by top 30 most abundance clonotypes. The peptide vaccine dose (in µg) used to induce Ag-specific T cells is shown below the graph. C, Length distribution of observed CDR3 amino acid sequences within each sample. Lines represent mean proportion within each group. Total TCR transcripts (D), Gini coefficient (E), number of unique CDR3 clonotypes (F), and clonality estimates (G) of Ag-specific repertoires. H, Differentially abundant clonotypes observed between groups. Datapoints represent individual values for each mouse. Statistical comparisons are determined by Student t test. Error bars show mean ± SD.