EMBO/sd-character-level-ner huggingface dataset:
Otoferlin dual-AAV injection at P6-7 partially restores synaptic function in Otof-/- IHCs.Figure 2High magnification views of IHCs immunolabeled for otoferlin and synaptic ribbons (CtBP2) from wild‑type (B6: P27, CD1B6F1: P29), dual-AAV injected CD1B6F1 Otof-/- (dual-AAV-TS: P26, dual-AAV-Hyb: P28), and their contralateral non-injected ears. (*) Transduced cells. Maximum intensity projections of optical confocal sections. Scale bars: 5 µm Synaptic ribbon numbers quantified from IHCs in apical cochlear turns of wild‑type (B6: n=48 IHCs, CD1B6F1: n=108 IHCs), transduced Otof-/- (dual‑AAV‑TS: n=59 IHCs, dual-AAV-Hyb: n=37 IHCs), and non-transduced Otof-/- IHCs from injected (-AAV injected ear, n=65 IHCs) and contralateral non-injected (-AAV non-injected ear, n=46 IHCs) ears (P25-29) IHC synapses labelled with CtBP2 and the postsynaptic marker Shank1a in B6 wild-type and Otof-/- P6 and P14 organs of Corti. Maximum intensity projections of optical confocal sections. Scale bars: 5 µm. D Synapse numbers quantified from IHCs in apical cochlear turns (C) of B6 wild-type (P6: n= 53 IHCs; P14: n=73 IHCs) and B6 Otof-/- (P6: n=62 IHCs; P14: n=65 IHCs) mice at two different developmental stages (P6 and P14) Ca2+‑current-voltage relationship of control CD1B6F1 wild‑type (n=6 IHCs), dual-AAV-TS transduced (n=8 IHCs), and non‑transduced CD1B6F1 Otof-/- (n=10 IHCs) IHCs (P14‑18) Representative Ca2+‑currents (Ica) and IHC plasma membrane capacitance increments (ΔCm) of a wild‑type control, transduced, and non‑transduced Otof-/- IHC in response to a 20 ms depolarization pulse at maximum Ca2+‑current potentials (typically -14 mV) Average exocytosis level measured as ΔCm (G) and corresponding Ca2+‑current integrals (QCa2+) (H) in wild‑type (CD1B6F1: n=6 IHCs; B6: n=11 IHCs (B6 data replotted from Strenzke et al, 2016)), dual-AAV-TS transduced Otof-/- (n=8 IHCs), and non‑transduced (n=11 IHCs) Otof-/- IHCs. Transduced IHCs that showed eGFP expression, but had almost no exocytosis are depicted as dashed lines (not included into the average)
ER stress regulates PIKfyve expression levels.Figure 2A : Time course of eIF2α and PERK phosphorylation in prion-inoculated C57BL/6 mice. T: terminal disease. B: PIKfyve/eIF2α and eIF2α(P)/eIF2α ratios assessed from Western blots as in A (n=3/group/time point). PIKfyve levels gradually decreased after 60 dpi whereas eIF2α became phosphorylated from 30 dpi until terminal disease. eIF2α phosphorylation and pPIKfyve depletion in POM1-treated tga20 COCS. **: p<0.01; ***: p<0.001; unpaired t-test; Error bars represent s.e.m. C: Treatment of COCS with POM1 reduces the levels of PIKfyve starting 48h post treatment. Each dot represents an individual experiment. Statistics: Unpaired t-test. **: p<0.01. D: Transient PIKfyve suppression in thapsigargin-treated Gt1 cells with recovery after 6 hours. PERK phosphorylation confirmed ER stress. E: Gt1 cells were inoculated with NBH or prions (60 dpi), pulsed with [35S]Met/Cys (20 min, 37 °C) and immediately lysed or chased without 35S (37 °C, ≤3 hours). For control, cells were treated with scrambled siRNA or siRNA against PIKfyve. Proteins were immunoprecipitated with antibodies to PIKfyve and subjected to SDS-PAGE and autoradiography (n=3 individual biological replicates). F: Autoradiographic signals expressed as percentage of the initial PIKfyve levels. Prion infection shortened the half-life of PIKfyve from 144 to 52 min. First-order kinetics was assumed. Error bars represent s.e.m and were calculated based on three individual biological replicates. G: Left: partially restored PIKfyve levels in prion-infected tga20 mice treated with LIN5044 (LCP) or vehicle. Right: quantification of the western blot. Each dot represents an individual biological replicate. *: p0.05; unpaired t-test Error bars represent s.e.m. H: Left, tga20 COCS were inoculated with RML or NBH, optionally treated with GSK2606414 starting at 21 dpi, and lysed at 35 dpi. Depletion of PIKfyve in prion-infected samples was largely rescued by GSK2606414 (GSK). Right, quantification of western blot. Each dot represents an individual biological replicate. Statistics: Unpaired t-test. **: p<0.01; unpaired t-test; Error bars represent s.e.m.
Figure 3Flow cytometric analysis of total B cells (A) as well as pro-B, pre-B, immature (imm) B and recirculating (recirc) B cells (B) from the bone marrow of wild-type (WT, gray), Prdm1ihCd2/+ (red) and Prdm1ihCd2/ihCd2 (white) mice at the age of 6-8 weeks Flow cytometric analysis of total B, FO B and MZ B cells from the spleens of wild-type, Prdm1ihCd2/+ and Prdm1ihCd2/ihCd2 mice Determination of cell death of ex vivo pro-B and pre-B cells from the bone marrow of Prdm1ihCd2/+ (red) and wild-type (gray) mice by flow cytometric analysis of the loss of plasma membrane asymmetry (F2N12S ratiometric dye staining) and the loss of cell membrane integrity (SYTOX staining). Representative flow cytometry plots (left) and the quantification of dead and apoptotic (apop) cells (right) are shown BrdU labeling of immature (imm) and FO B cells in the spleen of 3-month-old Prdm1ihCd2/+ mice (red dots) and control wild-type littermates (gray dots). The percentage of BrdU+ B cells was determined for each B cell type by flow cytometric analysis after 10 days of continuous BrdU labeling (day 10, white bar) or after a subsequent 15-day chase period (day 25; hatched bar) without BrdU in the drinking wate BrdU labeling of immature (imm) and FO B cells in the spleen of 3-month-old Prdm1ihCd2/+ mice (red dots) and control wild-type littermates (gray dots) A diagram (below) indicates the design of the BrdU labeling and chase experiments. Flow cytometric data obtained with FO B cells from one mouse of each genotype are shown in (F) The percentage of BrdU+ FO B cells is shown above the indicated gates (F)
Figure 2(A,B) Time course of representative binding/unbinding events in EAAT1 and GltPh simulations, shown as the distance of the closest K+ to the respective site. (C-E) Kinetic models of K+-binding steps for (C) EAAT1 and (D,E) GltPh (node radius, estimated equilibrium occupancy; lines, observed transitions; red and blue, two apo-K1 transition paths with the highest reactive flux).
Figure 8A, Peroxiredoxin levels in peroxiredoxin 1 and 2 double knockout cells (PRDX1/2 DKO). Lysates of the indicated cell lines grown in glucose-containing medium were analysed by immunoblot B. Peroxiredoxin levels in peroxiredoxin 1 and 2 double knockout cells (PRDX1/2 DKO). quantitative proteomics (B).The right subpanel in (B) lists all detected but not altered proteins belonging to cellular antioxidative systems. C. HyPer7 steady state ratios of the indicated cell lines. HyPer7 probes were targeted to the indicated compartments of the respective cell lines. Cells were grown in glucose-containing medium. In the boxplot, the central band is the median. The lower and upper hinges correspond to the first and third quartiles. The upper whisker extends from the hinge to the largest value no further than 1.5 times the inter-quartile range, whereas the lower whisker extends from the hinge to the smallest value no further than 1.5 times the inter-quartile range Data information: Statistical analysis was only performed to compare the PRDX1/2 DKO with the wildtype. D. Response of HyPer7 probes targeted to indicated compartments to treatment with 25 µM of exogenous H2O2. Indicated cell lines (black, wild type; red, PRDX1/2 DKO) were grown in glucose-containing medium. Solid line represents average, points colored in the lighter version of the respective color are the corresponding ratios measured in individual cells. Data information: Statistical analysis was only performed to compare the PRDX1/2 DKO with the wildtype. As most of the data were not normal distributed, instead of a t-test, a Wilcoxon/Mann-Whitney-U-test was performed and samples were compared in pairs. *, p ≤ 0.05, **, p ≤ 0.01, ***, p ≤ 0.001 E. Response of HyPer7 probes targeted to indicated compartments to treatment of mtDAO-expressing cell lines with 4 mM D-Ala (black, wild type; red, PRDX1/2 DKO; cell generated with the piggyBAC system). Cells were grown in glucose-containing medium. Solid line represents average, points colored in the lighter version of the respective color are the corresponding ratios measured in individual cells. Data information: Statistical analysis was only performed to compare the PRDX1/2 DKO with the wildtype. As most of the data were not normal distributed, instead of a t-test, a Wilcoxon/Mann-Whitney-U-test was performed and samples were compared in pairs. *, p ≤ 0.05, **, p ≤ 0.01, ***, p ≤ 0.001 F. Model. See discussion for details
Figure 3a. TCR repertoire clonality in inflammatory pseudotumor lesions (2015-2019) and 2019 CSF as analyzed by VDJ sequencing. Clonotype abundance represented by angular diameter. b. TCR repertoire distribution assessed as clonal space homeostasis, i.e., proportion of the repertoire occupied by the clones of a given size. c. Rarefaction analysis indicating estimated diversity of clonotype richness in inflammatory pseudotumor lesions (2015-2019). Sample size indicated on the x-axis. d. TCR sequence overlap between inflammatory pseudotumor lesions (2015-2019) measured by Morisita's overlap index. e. Shared (absolute number) and unique (proportion) clonotypes between inflammatory pseudotumor lesions (2015-2019). f. K-mer statistics computation and resulting motif probability matrix depicting occurrences of amino acids in each position of all computed CDR3 beta chains. g. Longitudinal tracking of most abundant clonotypes in inflammatory pseudotumor lesions (2015-2019). CDR3 amino acid sequence and v-chain usage for each clonotype annotated in the legend. Private clones are limited to each timepoint. Shared clones are found at all timepoints. h. Automated CDR3 beta chain high-resolution modeling of dominant T cell clone CASSQEYSPYEQY.
Sugarbabe is a downstream effector of ERK7 and PWP1A, B.Figure 7A, B. Expression of sugarbabe is downregulated in fat bodies upon knockdown of PWP1 (A) and ectopic expression of ERK7 (B) (N=3 replicates of 10 fat bodies/replicate for each genotype). C. ERK71 mutants fail to maximally downregulate sugarbabe expression upon fasting (N=4 replicates of 10 larvae/replicate for each genotype). D. Representative immunofluorescent images of fat bodies in a genetic epistasis experiment using ERK71 and sug mutants (sug17Δ/Df(2R) Exel7123) with LipidTOX and DAPI staining to visualize lipid droplets and nucleus, respectively. Scale bar: 50 µm. E. sug mutant (sug17Δ/Df(2R) Exel7123) suppresses increased lipid droplet number observed in ERK71 mutants (N>20). F. Ectopic sugarbabe expression in the fat body partially rescues TAG levels in the CG>ERK7 larvae (N=4 replicates of ≥10 larvae/replicate for each genotype). G. sug mutant (sug17Δ/Df(2R) Exel7123) suppresses increased nuclear size phenotype of ERK71 mutant fat body cells (N ≥25 cells per genotype). Ectopic Sugarbabe expression in the fat body partially restores the pupariation rate (H) (p<0.001 for the rescue, analyzed by Log-rank test, N=4 replicates of 30 larvae/replicate for each genotype) of CG>ERK7 larvae I. Ectopic Sugarbabe expression in the fat body partially restores the pupal volume of CG>ERK7 larvae (I) (N=4 replicates of 10 pupae/replicate for each genotype). J. Addition of 20% sucrose improves the growth rate and survival of CG>ERK7 larvae. Pupariation kinetics of CG>ERK7 larvae on 20% yeast or 20% yeast + 20% sucrose diet (p<0.0001 for the rescue, analyzed by Log-rank test, N=4 replicates of 30 larvae/replicate for each genotype and diet).
Figure 1Schematic representation of Terminator 5′-phosphate-dependent exonuclease (TEX) treatment of immunoprecipitated RNA. TEX digestion stops at the antibody-tethered nucleotide, while TEX eliminates the non-specific RNA and linkers remaining on the beads. Green circle represents m6A nucleotide. Distribution of RBFOX2-eCLIP reads and RBFOX2-tRIP reads on chr10: 75,544,000-75,635,000 (GRCh37/hg19). Distributions of FUS-HITS-CLIP reads, FUS-tRIP reads, and ContAb-tRIP reads on chr15: 26,165,000-28,520,000 (GRCm38/mm10). The number of N2A cells is indicated. Correlation of read densities constituting MACS-defined peaks between RBFOX2-tRIP and RBFOX2-eCLIP. Scatter plot indicates RPM of tRIP-seq and eCLIP constituting each peak. Correlation in read densities constituting MACS-defined peaks between FUS-tRIP and FUS-HITS-CLIP. Scatter plot indicates RPM of tRIP-seq and HITS-CLIP constituting each peak. The RBFOX2 motif identified by MEME in the peaks of RBFOX2-tRIP. The FUS motif identified by MEME in the peaks of FUS-tRIP of 1 × 108 cells. Distributions of FUS-tRIP reads (upper panel) and ContAb-tRIP reads (lower panel) mapped to the relative positions of all coding genes in mouse. The ngs.plot tool [58] was used to calculate the average RPM for a gene structure. The average RPM at each position was normalized based on the total RPM mapped to each gene. The standard error of normalized RPM is shown as a semi-transparent shade around the average curve. TSS, transcriptional start site; TTS, transcriptional termination site.
Autophagy is impaired in hepatocyte-specific ASK1-knockout mice (A) Shown is one representative Western blots from two independent experiments and quantification of respective protein levels in mice fed a HFD for 20 weeks (ASK1F/F n=8 mice; ASK1Δhep n=8 mice).Figure 4Shown is one representative Western blots from two independent experiments and quantification of respective protein levels in mice fed a HFD for 20 weeks (ASK1F/F n=8 mice; ASK1Δhep n=8 mice). Representative immunofluorescence images of liver sections stained for LC3-II punctate (red) and nuclei (DAPI, blue) in mice. Scale bar represents 100 µm.
Red1 localizes to cleavage bodies in mitotically dividing cells and cooperates with polyadenylation factors to hyperadenylate meiotic mRNAs.Figure 4(A) Red1 dots co‐localize with cleavage factor, Pcf11; the canonical poly(A) polymerase, Pla1; a nuclear exosome subunit, Rrp6; and a nuclear poly(A)‐binding protein, Pab2. Microscopic images of strains expressing both Red1‐tdTomato and either Pcf11‐, Pla1‐, Rrp6‐, or Pab2‐GFP are shown. Bars, 2 μm. (B) Red1 co‐immunoprecipitates with Pla1 and Rrp6. Total cell extracts prepared from strains expressing both Red1‐FLAG and Rrp6/Pla1‐myc were subjected to immunoprecipitation, followed by western blotting. (C) Meiotic mRNAs in red1Δ do not have highly elongated poly(A) tails. Northern blotting was performed to analyse rec8+ and spo5+ mRNAs in wild type (WT), red1Δ, rrp6Δ, and red1Δrrp6Δ. The same blots were stained with methylene blue to visualize ribosomal RNAs (rRNA), which serve as a loading control.
Figure 1A Quantification and representative immunoblots of lamin B1 in the striatum (8w, 12w and 20w; N = 6 for both genotypes; 30w; N = 4 and 5 for WT and R6/1 mice, respectively), hippocampus (8w; N = 8 and 6 for WT and R6/1 mice, respectively; 12w; N = 7 and 6 for WT and R6/1 mice, respectively; 20w N = 5 and 6 for WT and R6/1 mice, respectively; 30w; N = 8 and 5 for WT and R6/1 mice, respectively) and cortex (8w; N = 5 and 4 for WT and R6/1 mice, respectively; 12w and 20w; N = 6 for both genotypes; 30w; N = 6 and 5 for WT and R6/1 mice, respectively). Data information: Data is expressed as a percentage of controls. Each point corresponds to the value from an individual sample. Bars represent the mean ± S.E.M. *P < 0.05, **P < 0.01 and ***P < 0.001 as compared with WT mice (two-tailed unpaired Student's t test). Tubulin was used as loading control. Exact P values are reported in Appendix Table S3. B Quantification and representative immunoblots of lamin B2 in the striatum (8w, 12w and 20w; N = 6 for both genotypes, 30w; N = 5 for both genotypes), hippocampus (8w; N = 8 and 6 for WT and R6/1 mice, respectively; 12w; N = 6 and 7 for WT and R6/1 mice, respectively; 20w; N = 5 for both genotypes; 30w; N = 8 and 5 for WT and R6/1 mice, respectively) and cortex (8w and 12w; N = 6 for both genotypes; 20w; N = 5 for both genotypes; 30w; N = 8 and 5 for WT and R6/1 mice, respectively). Data information: Data is expressed as a percentage of controls. Each point corresponds to the value from an individual sample. Bars represent the mean ± S.E.M. *P < 0.05, **P < 0.01 and ***P < 0.001 as compared with WT mice (two-tailed unpaired Student's t test). Tubulin was used as loading control. Exact P values are reported in Appendix Table S3.
Figure 4A, C, E, G, I Fat bodies dissected from 4 hour starved animals expressing mCherry-Atg8a in all cells, and (A) sec5IR, (C) sec15IR, (E) sec3IR, (G) sec8IR, (I) exo84IR specifically in GFP-marked clone cells (yw, hsflp/w; pmCherry-Atg8a/+; act< FRT, cd2, FRT> GAL4, UAS-GFP/+) analyzed for mCherry-Atg8a puncta. Scale bars represent 20µm.B, D, F, H, J Quantification of data from (A, C, E, G, I). Data are represented as means ± SEM; n≥ 10. Statistical significance was determined using a Student's t test.
Figure 4A. Schematic representation of predicted domain structure of SPATA2. The figure shows domain structure of full-length SPATA2, and SPATA2 deletion mutants that were used for MS-based interaction analysis (see Figure 4B). B. The predicted PUB domain in the N-terminus of SPATA2 interacts with CYLD, but not with LUBAC or other TNF-RSC components. GFP-SPATA2 and the indicated deletion mutants were purified from A549 cells that were stimulated with TNF-α for 15 minutes, and the enriched proteins were quantified using SILAC-based MS. C. Validation of SPATA2 N-terminus interaction with CYLD. GFP-SPATA2 and the indicated deletion mutants were purified from A549 cells as described in Figure 5A, and association of CYLD was analyzed by immunoblotting. The lower panels show controls. D. The USP domain of CYLD interacts with SPATA2. STREP-FLAG-CYLD and the indicated deletion mutants were purified from A549 cells using Step-Tactin sepharose and blotted with SPATA2 antibody. The lower panels show controls. E. The C-terminus of SPATA2 interacts with HOIP. A549 cells were transiently transfected with GFP-tagged SPATA2, SPATA2 D200 and SPATA2 200*, and SPATA2 and the mutants were immunoprecipitated using GFP-trap beads. The immunoprecipitates were immunoblotted with HOIP, CYLD and GFP (HOIP) antibodies. The lower panels show input control, and the * shows unspecific band.
Figure 4(A) Heatmap indicating the normalized forward versus inverted reverse experiment enrichments on a log2 scale of quantitative proteomics upon pulldown of Flag-tagged Ythdf in S2R+ cells. The threshold was set to a 1-fold enrichment. Fmr1 is co-purified with Ythdf. Known Fmr1 interacting proteins are highlighted in bold with asterisk. (B) Co-immunoprecipitation experiment in S2R+ cells co-expressing FlagMyc-tagged Ythdf and Myc-tagged Fmr1. Ythdf was used as a bait via its Flag tag. The lysate was treated with Benzonase as indicated to remove interactions enabled by RNA. (C) GST pulldown of recombinant GST-Ythdf or GST alone mixed with recombinant 6xHis-NT-Fmr1. 2 µM of recombinant proteins were using during the pulldown. The pulldown of the proteins was analyzed by CBB staining. (D) Overlap of miCLIP dataset generated in S2R+ cells defining m6A modified transcripts with Fmr1 and Ythdf mRNA targets determined by TRIBE in S2R+ cells. The indicated P-value was determined by hypergeometric test.
PELP1 is modified by SUMO in a SENP3‐controlled process.Figure 5(A) HA‐PELP1 was expressed alone or in combination with SUMO1 or SUMO2 in the presence or absence of SV5‐tagged SENP3 in HeLa cells. Expression of the respective proteins was verified by western blotting. Detection of β‐tubulin served as loading control. (B) SUMO modification of endogenous PELP1 was monitored by anti‐PELP1 immunoblotting in HeLa cells transfected with siRNAs as indicated. Depletion of the respective proteins was verified by western blotting. (C) HeLa cells were transfected with siRNAs and plasmids as indicated. His‐SUMO conjugates were recovered on Ni‐NTA beads and subjected to western blotting using anti‐PELP1 antibodies. (D) HA‐PELP1 or HA‐PELP1K826R was expressed in the presence of SUMO1 or SUMO2 in HeLa cells. Expression of the respective proteins was analysed by western blotting. Detection of β‐tubulin served as loading control. (E, F) Flag‐p14ARF and SUMO were expressed in HeLa cells and SUMO modification of ectopically expressed HA‐PELP1 or endogenous PELP1 was monitored by immunoblotting with anti‐HA or anti‐PELP1 antibodies as indicated. Arrowheads indicate SUMO-PELP1 conjugates.
Inhibition of S6 kinase phosphorylation by Autophagy‐specific gene 1.Figure 3(A,B) HEK 293T cells transfected with denoted plasmids were (A) deprived of nutrients for 90 min, and then replenished with DMEM for 30 min or (B) stimulated with 50 ng/ml epidermal growth factor (EGF) for 30 min. The S6K immune complexes were analysed by immunoblot using appropriate antibodies (top three panels). Flag (ATG1α) blots were completed from the same cell lysates (bottom panel). (C) HEK 293T cells transfected with control siRNA, ATG1α siRNA or ATG1β siRNA were deprived of nutrients for 90 min, and then the phosphorylation and protein levels of endogenous S6K (top two panels) and S6 (bottom two panels) were determined. (D) HEK 293T cells transfected with control siRNA or ATG1α siRNA were deprived of nutrients for 90 min, and then replenished with DMEM for 30 min. The phosphorylation and protein levels of endogenous S6K were examined. For quantification, the levels of S6K Thr 389 phosphorylation were measured using Adobe Photoshop, and normalized to the S6K protein levels. Results are expressed as a fold change compared with the first indicated sample. (E) HEK293T cells were transfected with control siRNA or ATG1α siRNA with pEGFP‐C1 (a transfection marker, green), and nutrient‐starved for 90 min, fixed and stained with phosphospecific S6 antibody (red). ATG1, Autophagy‐specific gene 1; HA, haemagglutinin; KI, ATG1α Lys 46 Ile (kinase‐dead) mutant; siRNA, short interfering RNA; S6K, S6 kinase; WT, wild type.
Figure 6A Flow cytometry figures of CAF-1 body rescue experiment. First figure: Wildtype J-Lat 10.6. Second figure: CHAF1A knock-out J-Lat 10.6. Third figure: CHAF1A-KO J-Lat 10.6 was re-introduced with wildtype CHAF1A. Fourth figure: CHAF1A-KO J-Lat 10,6 was re-introduced with dLLPS-CHAF1A which harbored five mutations. The percentages of GFP-positive cells were labeled in the top right corner of each figure. B The statistical histogram of the results in (A). C Rescue experiment in heterogeneous Jurkat latency model which were performed as in (A). D, E The enrichment of H3K9me3 and H3K27me3 on HIV-1 LTR in different CHAF1A status. The reactivation efficiency in each group was indicated by the percentage of GFP-positive cells.
Figure 1Human monocytic (THP-1) cells were induced to acquire an M2-like phenotype (see Methods) and treated with different concentrations of TLR7-54 or FA-TLR7-54 for either 48h In the latter case, after the 2h incubation, culture medium was replaced with drug-free medium and incubation was continued for 46h. All treatment groups were then analyzed by qPCR for gene expression (A) Changes in mRNA levels of indicated profibrotic macrophage markers by induced by different concentrations of TLR7-54 and FA-TLR7-54 (n=3, technical replicates). Human monocytic (THP-1) cells were induced to acquire an M2-like phenotype (see Methods) and treated with different concentrations of TLR7-54 or FA-TLR7-54 for either 48h or 2h In the latter case, after the 2h incubation, culture medium was replaced with drug-free medium and incubation was continued for 46h. cell supernatants were analyzed for secreted cytokines by ELISA. (B-C) Changes in CCL18 and IL-1β in the culture media induced upon treatment with TLR7-54 and FA-TLR7-54 for the treatment regimens (n=3, technical replicates).