PC4 knockdown inhibits liver cancer cell growth in vitro and in vivo. Relative to Fig. 5. (A) qPCR and western blot showing PC4 protein and mRNA level of Huh7 and HepG2 cells stably expressing control shRNA (NC), human PC4-targeting shRNA (shPC4-1 and shPC4-2), or shRNA with PC4 expression (shPC4+PC4; the sequence for PC4 induction is optimized and resistant to shRNA). β-actin, loading control. Data were generated from n = 3 biological replicates. (B) Colony assay performed in Huh7 and HepG2 cells after PC4 stable knockdown. Data were generated from n = 3 biological replicates. (C) Soft agar assay performed in Huh7 and HepG2 cells with PC4 stable knockdown. Scale bar = 300 μm. Data are generated from n = 4 biological replicates. (D) EdU immunofluorescent staining performed in Huh7 and HepG2 cells with PC4 stable knockdown. Scale bar = 75 μm. Data were generated from n = 3 biological replicates. (E) Tumor weight in the nude mice subcutaneously injected with Huh7 cell stably expressing NC, shPC4-1, shPC4-2, or shPC4+PC4. Data were generated from n = 5 biological replicates. (F) Representative images of PC4, Ki67, and CCND1 immunostaining in tumor tissue of indicated xenograft mice. Scale bar = 75 μm. (G) Histograms show the cell cycle profiles in NC or PC4-knockdown Huh7 cells treated with double-thymidine block, followed by releasing for an indicated period of time. Data are generated from n = 3 biological replicates. All the data were shown as means ± SD (one-way ANOVA test); error bars represent SD. **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are available for this figure: SourceData FS3.