Fig. 1: Identification of USP26 as a potential regulator in HCC cells through CRISPR-based DUB library screening. | Nature Communications

Fig. 1: Identification of USP26 as a potential regulator in HCC cells through CRISPR-based DUB library screening.

From: USP26 as a hepatitis B virus-induced deubiquitinase primes hepatocellular carcinogenesis by epigenetic remodeling

Fig. 1

a Schematic strategy for CRISPR-based DUB library screening. A total of 348 sgRNAs targeting 87 DUBs (4 sgRNAs/DUB) were transfected into the HCCLM3 cell line, which was subsequently screened. Left: Dot plots showing the rank of DUBs from 1 (lowest) to 87 (highest) based on colony formation assay (b) and EdU staining assay (c) of sg-DUBs HCCLM3 cells, which were measured and calculated by log2fold change. The top DUBs with inhibitory effects on cell growth were listed as indicated. Right: The percentage of significantly changed DUBs in each group (p < 0.01, log2 fold change < −0.58, down group; p < 0.01, log2fold change > 0.26, up group; p > 0.05, −0.58 <log2 fold change < 0.26, no difference group (No-diff)). d Venn diagram showing the overlapping DUBs that inhibited cell survival in the two screening assays described in b and c. e, f Western blot analysis and colony formation assay in HCCLM3 cells with or without CRISPR/Cas9-mediated USP26 knockout. g Flow cytometry analysis showing the percentages of cells in S phase in the sg-Control and USP26 knockout groups. h. Representative immunoblots showing USP26 protein levels in tumors and paired normal tissues from HBV-positive (HBV+) HCC patients. N, normal tissue; T, tumor tissue. Quantification of relative USP26 protein levels in tumors and paired normal tissues from HBV+ HCC patients (i) and HBV-negative (HBV−) HCC patients (j). k Kaplan-Meier survival analysis of HCC patients with high or low USP26 expression. The data were obtained from the Kaplan-Meier plotter database. Each graph is presented as the mean ± SEM. b, c n =  3 biologically independent experiments; (e–h) n =  3 biologically independent experiments; n = 42 (i) and n = 8 (j) paired tumor and normal samples. Statistical significance was calculated by (b–d) one-way ANOVA; (e, g) two-tailed unpaired t-test; (i, j) two-tailed paired t-test; (k) log-rank test. Source data are provided as a Source Data file.

Back to article page