Fig. 4

Knockdown of ATIC expression suppresses growth of HCC cells. a, LO2 cells were infected with indicated shRNA for 72 h. The efficiency of ATIC knockdown was determined by western blotting. b, Representative images of LO2 cells 5 days after transfection with indicated shRNAs. c, LO2 cells were infected with indicated shRNA for 72 h. Cell apoptosis are analyzed by flow cytometry. The apoptosis percentage was plotted. N = 3, NS, no significant. d, Representative images of HCC cells 5 days after transfection with indicated shRNAs. e, Cell numbers at indicated days after transfection with indicated shRNAs. N = 4, **, p < 0.005, ***, p < 0.001, compared to shRNA 1 and shRNA 4 groups. f, Cells were transfected with indicated shRNAs for 72 h followed by Hoechst and EdU labeling analysis proliferation. Representative images were shown. EdU-labeled (pink) indicated proliferation cells. Bars, 10 μm. g, the proliferation rate was plotted. N = 4, **, p < 0.005. h, Cells at 72 h post transfection with indicated shRNAs. Shown are representative flow cytometry plots. i, the apoptosis percentage was plotted. N = 3, **, p < 0.005. j, Cells were transfected with indicated shRNAs for 72 h, followed by western blot analysis of caspase 3 activation. k, Cells were transfected with indicated shRNAs. 24 h after transfection, 1500 cells were seeded in 12-well plates for 10 days followed by staining and quantification. Representative images were shown. l, colony number was plotted. N = 3, **, p < 0.005. m, relative colony size was plotted. The colony size in shScr group was set as 1. **, p < 0.005