EMBO/sd-character-level-ner huggingface dataset:
D. Phosphorylation status of Ser127 or Ser128 determines subcellular localization of YAP. Immunofluorescence analysis was performed in HeLa cells transfected with EGFP-YAP, EGFP-YAP-S127A, EGFP-YAP-S128A, or EGFP-YAP-S128D at high cell density. Figure shows a representative of multiple areas. Nuclei were stained with Hoechst 33342. Scale bars: 20 μm.
(B-C) miR-515-5p inhibits random (B) and directed (C) cell migration. The indicated cell lines were transfected with miR-515-5p for 48 h before time-lapse imaging was performed for 18 h (A) or transwell migration assays were performed for 9 h (C). (B-left panel) Plots show overlays of representative trajectories travelled. (B-right panel) The distance of migration was quantified and represented as the mean ± SEM of values normalised to the respective control condition. (C-left panel) Representative field of view from the bottom of the transwell chambers. Scale bar is 20 µm. (C-right panel) Data represent percentage of the average number migratory cells per field (n=5 fields per condition). Data are mean of three experiments ± SEM. (B-C) P values were calculated by t-test between miRNA (515-5p) values and their respective non-targeting control miR (NC) values (***, P<0.001). All data shown are representative of experiments performed at least in triplicate.
D Immunoprecipitation assay showing the interaction of DDX6 or DDX6 Mut1 with GFP-NOT1 in HEK293T cells. Inputs (1% for the HA-tagged proteins and 3% for the GFP-tagged proteins) and immunoprecipitates (20% and 15%, respectively) were analyzed by western blotting using anti-HA and anti-GFP antibodies. Endogenous CNOT2 and CNOT3 (input 0.5% and IP 10%) were detected using specific antibodies (Appendix Table S4).
(E) Contribution of pyruvate and glucose-derived carbon to TCA cyle metabolites in Flp-In 293 HA-GFP or Flp-In 293 HA-IKKε cells treated with doxycycline (50 ng/ml, 16 hours). (n=5 technical replicates). metabolite levels were normalised to the internal standard HEPES.
(c) Accumulation of Beclin 1 and APG5-APG12 complex in Bcl-2-transfected MEFs after etoposide treatment. WT MEFs were transfected with the indicated plasmids, and were incubated without (NT) or with 20 μM etoposide for 24 h.
F. Representative images of SCNT embryos at 115 h after injection of different mRNA as indicated. Scale bar, 50 μm
Decreased levels of synaptic IL1RAPL1 protein in the Ptprd-/- brain (P21-27), as revealed by immunoblotting of crude synaptosomal (P2), synaptic plasma membrane (SPM), and PSD (PSD II) fractions. Analysis of immunoblots reveal significant decrease of IL1RAPL1 in the SPM and PSD fractions of Ptprd-/- brain samples. (n = 3 mice [WT and KO] for each fraction [P2, SPM, and PSD], mean ± SEM, **p<0.01, Student's t-test).
(F,G) miniature excitatory postsynaptic currents (mEPSCs). (F) example traces of mEPSCs in WT and Clcn3-/- neurons (G) No significant difference between Clcn3unc/unc/Clcn4-/- and Clcn3+/+/Clcn4-/- mEPSCs in mean amplitudes (p > 0.9, t test) or in the distribution of amplitudes (Clcn4-/-: 12 cells, 2 mice; Clcn3unc/unc/Clcn4-/-: 8 cells, 2 mice).
(C) Lysates of MEFs that had been transfected with control or Rab12 siRNA were analysed by immunoblotting with the antibodies indicated.
C. Schematic model of impact of restoration of DCC-induced cell death by using a netrin-1 targeting antibody.
(D) Immunoprecipitation of Ago3 with Flag-tagged DDX43 WT and its ∆KH mutant lacking KH domain. Ago3 and DDX43 were detected by western blotting using anti-Ago3 and anti-Flag-antibodies, respectively. Flag-EGFP was used as a negative control. The ∆KH mutant maintains affinity to Ago3. IgG h.c.: IgG heavy chain.
A, B Quantification of different types of spermatozoa in fertile men with or without USP26 mutations. The spermatozoa from fertile men without USP26 mutations (n = 10 independent experiments); fertile men contained the USP26 c.463C>T mutation (The sample was analyzed in three repeated experiments), the USP26 c.125T>C mutation (n = 2 independent experiments, one sample was analyzed in two repeated experiments), the USP26 c.1044T>A mutation (n = 5 independent experiments), and the USP26 c.370_371insACA/494T>C/1423C>T mutation (n = 10 independent experiments) were collected to perform Chr X and Chr Y FISH. P-values are shown for comparison to fertile men without USP26 mutations.
(G) Pulldown assay using GST-SUMO IκBα protein and chromatin lysates from 2-month-old WT mice villi.
A-H Intracellular localization of LAMP1-positive EL, total (HA) and polymeric (2C1) ATZ in WT MEF exposed to BafA1 for 12 h (A); in Atg4BKO MEF (B); in Atg7KO MEF (C); in Fip200KO MEF (D); in Ulk1/2 double-KO MEF (E); in Atg13KO MEF (F); in Atg9KO MEF (G); in RubiconKO MEF (H). Scale bars: 10 μm. I Quantifications of A-H (n=10, 9, 9, 10, 7, 15, 9, 12 cells). One-way ANOVA and Dunnett's multiple comparisons test, ns P>0.05, **** P<0.0001.
A. Experimental design and representative traces for analysis of synaptic transmission in retinas from wild-type (black) and RIBEYE KO mice (red). Presynaptic bipolar neurons were depolarized in voltage-clamp mode from -70 mV to -10 mV for 0.5 s as indicated on top to open Ca2+-channels and trigger release. Presynaptic Ca2+-currents and postsynaptic EPSCs were measured simultaneously as indicated in the traces; dotted box displays an expansion of the initial Ca2+-currents and EPSCs.B. RIBEYE KO does not alter presynaptic Ca2+-currents. Summary graphs show average peak Ca2+-currents and Ca2+-current.C. RIBEYE KO severely impairs EPSCs triggered by presynaptic depolarization. Plot shows integrated EPSC charge as a function of time; both initial and sustained neurotransmitter release are impaired.D. RIBEYE KO strongly reduces initial synchronous release induced by presynaptic depolarization as indicated by the summary graph of the peak EPSC amplitude.E. RIBEYE KO suppresses both the initial synchronous and the sustained phase of release induced by presynaptic depolarization. Summary graphs display integrated EPSC charges for the initial phase (0-50 ms) and the sustained phase (50-500 ms).F. RIBEYE KO does not alter the kinetics of EPSCs. Summary graphs depict the mean synaptic delays (left), rise times (center) and decay time constants (right) of EPSCs.
(B) The islets from the mice (A) were treated with 3.3 mM (LG) or 30 mM glucose (HG). After lysis, the lysates underwent immunoblotting analysis using the antibody against Beclin 1, LC3 , p62 and actin.
B. Representative flow cytometry plots showing gating strategy to identify and compare activated B cells (CD138+ Blimp1+), Pre-PB cells (CD138- Blimp1+) and plasma blast (PB) cells after four days of stimulation with LPS either in the presence of DOT1L inhibitor, Pinometostat or DMSO as a control (left panel) and statistical analyses of their relative numbers (right panel). Results represent the data from one experiment and numbers represent biological replicates for each treatment (n = 4).
H) Extension reflex as a function of time in another experiment Gbeys/ys with vehicle (n=8), or 144DG11 (n=9), or wt mice with vehicle (n=8). Data information: Two way ANOVA with repeated measures show that, throughout the period, treatment values were higher than vehicle (p<0.05) Significant difference (p<0.05, two-tailed t-tests) at specific time points is denoted by *. All error bars represent s.d.
(E) Peptide ELISAs detect anti-mouse TREM2 antibody binding to tiled stalk region peptides, full length stalk peptide or a truncated ADAM cleavage site peptide. The binding epitope of 4D9 antibody is located 12 amino acids N-terminal of the ADAM cleavage site at His 157.
(E) ChIP analysis performed in BY4741 wild-type, nat4Δ and Pste5-NAT4 strains grown under NCR and CR conditions. Chromatin was immunoprecipitated using antibodies against N-acH4 and Histone H4, and analyzed by qRT-PCR with the indicated primers. PNC1-P (Promoter), PNC1-S (Transcriptional Start), PNC1-E (End of gene). The enrichment from the antibody was normalized to Histone H4.
Colocalization of novel nSB proteins with HSATIII. THRAP3 (F), were stained with anti-THRAP3, HSATIII was visualized by RNA-FISH using a dig-labeled HSATIII ASO, and the nuclei were stained with DAPI. Scale bar: 10 μm.
A We co-transplanted barcoded WT HSCs with competitor WT, B cell deficient (uMT-/-), or B and T cell deficient (NSG or Rag2-/-γc-/-) HSCs into irradiated recipient mice. Peripheral blood was harvested from recipient mice at 6 to 8 months after transplantation and sorted into granulocytes (Gr), B, CD4 T, and CD8 T cells for population and clonal level analyses
(D,E) RNA affinity chromatography using the different RNA constructs depicted in (i), as described in A.
(B) Global editing levels in the 3 brain regions is based on the Alu editing index, representing the weighted average editing level across all expressed Alu sequences, Wilcoxon, corrected p*=0.038, 0.055 and 0.42 for Amygdala, MTG and SN. Wilcoxon test between the Alu editing index of control and PD samples.
(E) IR abundance measured by immunoblotting following 4 hours pre-incubation with the HDAC6 inhibitor (Tubastatin A) in differentiated adipocytes exposed to insulin (100nM) stimulation for 15 min (n=4). Data are presented as means + SEM; Student's unpaired t-test and ANOVA with Tukey or Dunnet's post hoc test: *p<0.05, **p<0.01, NS= no significance.
B TE families with tissue-specific, singleton TSD-bearing insertions detected in pooled gut or head DNA.
E HCT116 cells were treated with actinomycin D for the indicated times. Cells were fixed and immuno-stained as indicated. Immunofluorescence images were taken by confocal microscopy. Scale bar represents 10 μM.
F Confocal images showing the basal region of organ of Corti cultures treated with either culture medium alone or medium containing 10 µM CDDP for 1 day and immunolabeled for Myosin 7A (red) and p-Chk2 (green). Scale bar=15 µm.G Histograms of green fluorescent signal intensity from OHC and IHCnuclei of a CDDP-treated organ of Corti explant.H Quantification of Chk2-positive nuclei in both IHCs (red bars) and OHCs (blue bars) from control and CDDP-treated organ of Corti cultures (n=50 nuclei per condition and per time point). Data are expressed as mean ± SEM. Kruskal-Wallis test followed by post hoc Dunn's test (*P=0.045, ***P=0.0002, CDDP-exposed IHCsvs. control IHCs; CDDP-exposed OHCsvs. control OHCs).
(B) Cherry-STING, IFNβ-Luc, pRL-TK, cGAS-GFP (stimulated) or IRES-GFP (unstimulated) and indicated expression plasmids as shown in (A) were transiently expressed in 293T cells and a dual luciferase assay was performed. Data is combined from two out of three independent experiments and shown as mean ± SD. Data information: Student's t-test (unpaired, two-tailed), n.s. not significant, **p<0.01, ***p<0.001, ****p<0.0001
(F) Statistically significant directionality in the overlap of the synergistic genes with the Hox/Meis1_DN gene set (Dataset EV5B) including Prmt1, which has also been independently validated by RT-qPCR (right panel). See also Appendix figure S5
(F) BIR frequencies (Lys+ survivors among total cells) for the WT and indicated mutant strains are plotted on a logarithmic scale. Data represent the mean ± SD from 3 to 6 independent experiments. ns, non-significant difference, * P-value = 0.0134, Mann-Whitney unpaired t test.
(A) p53-dependent dephosphorylation of Ulk1 at Ser637. p53+/+ and p53−/−MEFs were treated with etoposide (10 µM), and the expression of each protein was examined by western blotting. Dead cells (%) indicates the population of apoptotic cells assessed by propidium iodide (PI) staining. α-Tubulin was used as a loading control. A semiquantitative analysis of protein expression is shown in Fig. EV1.
(C) Close up of protophloem sieve element strands expressing truncated MAKR5-CITRINE fusion proteins under control of the UBQ10 promoter.
D RT-qPCR analysis of TA-ERVs in U2OS and HeLa cells ectopically expressing TERT-WT or TERT-K626A. E RT-qPCR analysis of TA-ERVs in HeLa and HCT116 cells with TERT knockdown. F
Western blots and quantification (right panels, relative to actin) showing levels of pSTAT3 Y705 and total STAT3. Upper panel: Data for Dgcr8flox/Δ (control) and Dgcr8Δ/Δ cells 5-6 h following serum differentiation (n = 2). Light grey and dark grey bars represent the data for 2 experiments. Lower panel: Data for BakΔ/Δ;Baxflox/Δ;Dgcr8flox/Δ GPCs (control) and BakΔ/Δ;BaxΔ/Δ;Dgcr8Δ/Δ (TKO) cells during serum differentiation (n = 3). Quantification is shown on the right; error bars represent standard deviation.
Representative dual-channel kymographs (A) and quantitative analysis (B showing that overexpression of Snapin, but not Snapin-L99K, a Snapin mutant deficient in dynein motor binding, reduces mitophagic accumulation within AD axons incubated with CCCP.
F-G) Phagocytosis assay on E16 (F) and WL hemocytes (G) srp(hemo)-moesin-RFP. The beads (in green) are phagocytosed by the hemocytes (in red). The WL hemocytes show greater phagocytic capacity compared to the embryonic ones after 5 min of exposure. Full stacks are displayed and the scale bars represent 20 µm.
(B) Calcium responses in individual tumors cells were recorded in (upper table) control tumors (developing without T cells) or (lower table) after T cell transfer in tumor cells contacted by at least one CTL during the imaging period. Data is representative of three independent experiment. Data information: Data were pooled of four independent experiments. (*p<0.05, ***p<0.001, unpaired t‑test).
A. When an Anopheles mosquito feeds from an infected mammal, it takes up parasites with the bloodmeal. The parasites transform into motile ookinetes in the midgut. Myosin is essential for traversal of the midgut epithelium. Ookinetes transform into oocysts and develop into hundreds of sporozoites. Sporozoites egress from oocysts to float in the hemolymph. Active motility allows sporozoites to invade the salivary glands from where they are transmitted into the skin with the next bite. They traverse the skin and enter into a blood vessel. With the bloodstream they are transported until they reach the liver and actively invade a hepatocyte to develop into thousands of merozoites. These merozoites are released back into the bloodstream where they depend on myosin to actively invade red blood cells (RBCs) and multiply. Red and blue arrows indicate active and passive movements of the parasite, respectively.
(F) Electron micrographs ( × 10,000 magnification) of BAT depicting mitochondria from 4‐mo‐old Con and KO mice. m, mitochondria; LD, lipid droplet; n, nucleus.
D. Images show pole-to-pole tracking (white curve) of PRC1-GFP signal (top, merge; middle, GFP; bottom, scheme) from a HeLa cell expressing PRC1-GFP and mRFP-CENP-B. Images of the same spindle without (left) and with the tracked contour (right) are shown. The graph shows normalized pole-to-pole intensity profiles (legend as in C) of PRC1-GFP acquired in HeLa cells expressing PRC1-GFP and mRFP-CENP-B.
D. Overlay of cryoEM maps of the H3 nucleosome (green; PDB 6ESF) and the CENP-A nucleosome (red). Note shorter and moved density for DNA on the CENP-A nucleosome indicated by the arrow.
Figure 3: Infiltrating leukocyte subsets in human glioma. Paraffin sections of healthy human brain (n=3), low grade glioma (n=12), anaplastic astrocytoma (n=7) and glioblastoma (n=9) were stained for IBA1 (macrophages/microglia), CD15 (granulocytes) and CD3 (T-cells) (A). Quantification of leukocyte subsets in human glioma specimens (B). For statistical analysis Kruskall-Wallis (Dunn´s post test) was applied.* p < 0.05, ** p < 0.01; data are mean ± SEM
Survival probabilities of in which CheY was found to interact with FliM. Black line, a bi-exponential fit; dashed line, a fit of the fast decline process F, phosphorylated CheY(I95V)-Atto647 (ΔcheZ background; strain EW669). 82 cells were recorded with a total of 2414 trajectories.
(E) HeLa GFP-LC3 cells were transfected with indicated siRNAs for 48 hr and then infected with DsRed-labeled Salmonella for 1 hr. The fraction of GFP-positive intracellular Salmonella is shown. Data are mean ± SD and are representative of three independent experiments.
D Venn diagrams of up- (left) and down-regulated genes (right) for all three conditions (+/-1.5-FC and p<0.01). Heatmaps of genes only deregulated in MaxKO-cd19, MycKO-cd19, or DKO-cd19 mice are shown.
(D) The soft SGGFP or BCL9L-SGs- 4T1, MCF-7, B16 or MP-1 cells (100 cells) were injected into the mammary fat pads (4T1 and MCF-7) or subcutaneous tissue (B16 and MP-1) of NSG mice. Tumor formation was recorded. n = 10 or 12 mice as indicated.
Representative images of the H&E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (D), distribution of the fibers cross-section area (CSA) (E), percentage of myofibers with central nuclei (F) in GA muscle from WT and mGPDH-/- mice at day 7 post CTX injection
WT and LRRK2 KO RAW264.7 macrophages were infected with Ca for 60 min Galectin-3 and LC3B recruitment was analysed by immunofluorescence and high content imaging. Scale bar = 10 μm. Data show the mean ± SEM of two to three experiments. ns=non-significant, *p≤0.05 by Student's t-test.
The levels of ATP6V0D in the membrane fraction are shown in (D) after normalization against LAMP1 (n = 3 independent experiments, ns represents non-significant). Data information: Error bars represent ± standard deviation. Scale bar, 10 μm.
Ratios of IgG to IgM derived from ELISA values plotted against molar ratios of antigens for n=5/group. Mean ± SD, statistical significance was calculated by using Kruskal-Wallis test, *P < 0.05, ***P < 0.001.
(C) Model for the roles of Bur1, TORC1 and Sch9 for progression through G1 phase of the cell-cycle. Bur1 and TORC1 act in parallel to phosphorylate Sch9. We propose that similar to TORC1, Bur1 phosphorylates Sch9 on the vacuole membrane, or alternatively that Sch9 which is phosphorylated by TORC1 on the vacuole, translocates to the nucleus where it is further phosphorylated by Bur1. Then Sch9 acts in parallel with Cdc28/Cdk1-Cln3, in a process that results in the export of Whi5 from the nucleus and cell-cycle progression through early G1 phase.
(C) Single confocal sections of HeLa cells. CREST and CENPT were used to label the centromere region and associated KTs, respectively. Enlargements of boxed areas displaying both chromosomes aligned and not aligned at the metaphase plate are shown. (D) Average of the inter KT distance based on in each of which 10 chromosomes/cell and 10 cells/sample were counted. P values are obtained by Kruskal-Wallis test with Dunn's multiple comparison analysis. Note that both aligned and not aligned KTs have a shorter inter KT distance compared to aligned control KTs.
G Analysis of relative collagen type I content identified by Sirius Redstaining 7 days after MI. Student`s t-test; n = 4 mice for ZT5 and n = 5 for ZT13 MI; *P = 0.0175.
Western blotting of Myc-SV40-NLS-Egg WT in whole (Whole), cytoplasm/nucleoplasmic (Cyto/Np) and chromatin (Chromatin) fractions before (-) and after Flag-Wde co-expression. Anti-Myc and anti-Flag antibodies were used. Histone H3 was detected as a marker of chromatin fraction.
G. Heat map for select DEGs of the lungs from aged mice treated with IgG (in triplicate), high dose (αCD8Hi, in triplicate) or low dose (αCD8Lo, pooled n=3) of CD8 Ab as analyzed by Nanostring. Orange is relatively high expression and purple is relatively low expression.
(F) Western blot analysis of EVs isolated by UC from HCT116 cells in glutamine-replete medium, transduced with a Rab7 or non-targeting (NT) control shRNA knockdown construct. Bar charts show changes in putative exosome proteins in EVs isolated by ultracentrifugation, following normalisation to cell lysate protein. (F') Growth curves are for HCT116 recipient cells pre-treated with EVs isolated as in (F) or with vehicle (PBS).
, C) Sequential dissociation of EI24 and BiP from IRE1 elicits a two-phase activation of IRE1 in response to ER stress. Cells were treated with 4 μM tunicamycin (TM) for the indicated times. IRE1 phosphorylation was analyzed by immunoblotting. The curve displays the coordination of IRE1 phosphorylation with the dissociation of EI24 and BiP (C).
(g) Effects of wild-type CerS1 (−/+ tet) on ATP generation in the absence (control shRNA) or presence of shRNA-mediated knockdown of LC3B. Data shown are an average of at least three experiments ± s.d. (*P 0.05).
H, I. Wnt inhibition reduces the expression of HR and FA pathway genes in a RNF43-mutant pancreatic cancer patient-derived xenograft and AsPC-1 cells. (H) Pancreatic canc­er PDX with G371fs RNF43 mutation treated with vehicle or ETC-159 (30 mg/kg) for 21 days were analysed for changes in the expression of the indicated genes measured by qRT-PCR. Each data point represents an individual tumor. The horizontal lines represent mean of 3 tumors/group. (I) AsPC-1 cells were seeded in low adherence plates, treated with DMSO or ETC-159 (100 nM) for 72 hours and the expression of indicated genes was measured by qRT-PCR.
Impact of Ctcflos KD on Ucp1 expression is largely mediated via reduced Prdm16 transcription. (H) Relative Ucp1, (I) Ctcflos tr1 and (J) Prdm16 transcript levels in single and double KD (day1 diff) of Ctcflos tr1 (ASO 1) and Prdm16 (siPrdm16), nontargeting LNA Gapmer (light grey) and siRNA (dark grey) controls, assessed by qPCR, mean and individual values, one-way ANOVA (Šídák-test), n.s. p>0.05, *p<0.05, **p<0.01, ***p<0.001.
B MOAP-1 downregulates the p62 bodies in the resting HepG2 liver cancer cells. HepG2 were transfected with plasmid encoding Myc-MOAP-1. At the indicated time post-transfection, transfected cells were subjected to IF as in (A). Data information: nuclei were counterstained with DAPI (blue)
B Global average MSD curves of mEOS-Cdc42 in the strains indicated at the Pole (P) and Non-Pole (NP) of cells. Trajectories longer than 6 frames were analyzed. Number of trajectories analyzed: BEM1 (n = 11 cells: Non Pole: 1814 tracks; Pole: 706); bem1 bc-14E px (n = 10 cells: Non Pole: 1793 tracks; Pole: 908); bem1 bc-14E (n = 13 cells: Non Pole: 1714 tracks; Pole: 975). Error bars display s.e.m. C D coefficients of mEOS-Cdc42 in the strains indicated (in box-plots displaying the median (line), the 25-75 percentiles (box) and the mean (cross)), which were compared using a non-parametric, two-tailed Mann-Whitney rank sum test.
Quantification for the width of intestinal lumen in N2 exposed to E. coli and P. aeruginosa for 12 h. ns (not significant) P > 0.05 as determined by two-tailed unpaired t-test. n = 3. Error bars indicate SEM.
(L) Probability of unimodal distributions of death times calculated by Hartigan's dip significance (mean of three independent experiments ± standard deviation; two-sample t-test **P<0.01).
(E) The corresponding molecular signatures of PSD activity represented by the label distribution per molecular species between *PS and *PE as percentages of the total amount of label incorporated in the molecular species indicated.
(A) WT and Srcap-/- ISCs were conducted for transcriptome microarray, followed by Gene Set Enrichment Analysis (GSEA) analysis for PPARδ target genes.
G. Relative mouse weight (%) during treatment of GSK-LSD1 (n = 8) or vehicle-treated (n = 8) mice
(D) Structural comparison between the YAP-WW1/SMAD7-PY and STXBP4-WW/SMAD7-PY complexes. The identified 9 amino acid residues were indicated for both complexes.
E. Venn diagram shows the differentially expressed (DE) genes between adult Langerhans cells (LC) and adult F4/80hi or CD11bhi cells (functional connectivity |FC| > 1.5, FDR adjusted p-value < 0.05).
IB analyses of MT-HCT116 cells transfected with GFP-Mock, -BBR-WT, or -BBR-2E for 24 hr and then treated with 20 µM of KYA1797K for 24 hr
For all the co-staining experiments IFNγ-stimulated HFFs infected for 3 h with the Pru strain were used. For each staining at least 50 vacuoles were scored (n=3). On the right-hand side, a representative fluorescent image is shown for the Toxoplasma Pru strain. Scale bar is 20 µm. The yellow box inside each representative image is shown as inset pictures with higher magnification. The total number of coated vacuoles was set at 100% and the percentage of vacuoles positive for Ub, GABARAP was calculated.