Fig. 4.

Suppression of anchorage-independent proliferation by silencing of PKR or 4.1R in HCC cell lines. (A) HuH7 cells (top panel) and HepG2 cells (bottom panel) on culture dishes were transfected with control siRNA or two 4.1R siRNAs, followed by MTT assay. The black line, blue line, and yellow line show the results for control, 4.1R siRNA1, and 4.1R siRNA2, respectively. (B) Huh7 and HepG2 cells were transfected with control siRNA or two 4.1R siRNAs, followed by colony formation assay. Three representative images of colony formation on soft agar and their quantified colony area with control siRNA or two 4.1R siRNAs, respectively, in HuH7 (B, top panels) and HepG2 cells (B, bottom panels) are shown. (C) HuH7 cells (top panel) and HepG2 cells (bottom panel) on culture dishes were transfected with control siRNA or two PKR siRNAs, followed by MTT assay. The black line, blue line, and yellow line show the results for control, PKR siRNA1, and PKR siRNA2, respectively. (D) Huh7 and HepG2 cells were transfected with control siRNA or two PKR siRNAs, followed by colony formation assay. Three representative images of colony formation on soft agar and their quantified colony area with control siRNA or two PKR siRNAs, respectively, in HuH7 (D, top panels) and HepG2 cells (D, bottom panels) are shown. (E) Huh7 and HepG2 cells were transfected with the indicated siRNA and plasmid, respectively, followed by colony formation assay. Three representative images of colony formation on soft agar and their quantified colony area after transfection of the indicated siRNA and plasmid, respectively, in HuH7 (B, top panels) and HepG2 cells (B, bottom panels) are shown. The values indicate the mean ± SD values of four independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the control group by the two-sided Student’s t-test or Tukey’s test.