Fig. 2: ASF1a knockdown leads to growth arrest and cellular senescence in wild-type p53-carrying HCC and PCa cells.

a Western blot was performed for the validation of ASF1a siRNA efficiency in HepG2 and LNCaP cells. ASF1a si1 and ASF1a si2, two different ASF1A siRNAs, were used in this study. b The cell number of control (nc siRNA) and ASF1a knockdown (ASF1a si1/si2) groups in HepG2 and LNCaP cells 72 h after siRNA transfection. Cells in negative control groups were set as 100% (reference). The siASF1a-transfected groups showed a significant decrease in cell counts compared with control cells (data are presented as the mean ± SD of four independent experiments in HepG2 and three independent experiments in LNCaP, respectively; P values are shown in each panel). c Colony-formation assay of HepG2 and LNCaP cells. The clonogenic abilities of HepG2 and LNCaP cells were decreased upon ASF1a knockdown. Quantification is shown at the bottom (data are presented as the mean ± SD of three independent experiments in HepG2 and four independent experiments in LNCaP, respectively; P values are shown in each panel). Colonies in control groups were set as 100% (reference). d Cell cycles of HepG2 and LNCaP recorded by propidium iodide (PI) staining. Cell cycles in control groups are shown with red peaks and cell cycles in ASF1a knockdown groups are shown with blue peaks. Proportions of G0/G1, S, and G2/M phases are presented in pie charts (data presented are of three independent experiments in HepG2 and four independent experiments in LNCaP; P values are shown in each panel). e β-Gal staining of HepG2 and LNCaP cells. Senescent cells were stained with blue color (scale bar: 100 μm). f Quantification of β-gal staining-positive HepG2 and LNCaP cells (data are presented as the mean ± SD value of three independent experiments for HepG2 and LNCaP, respectively). HCC, hepatocellular carcinoma; PCa, prostate cancer