Fig. 3

IFI6 promotes cell proliferation, inhibits apoptosis and ameliorates oxidative stress in ESCC. a-b. Representative images (a) and statistical quantification (b) of EdU staining in ESCC cell lines transfected with IFI6-shRNA or ShControl lentivirus. EdU: red, Hoechst 33342: blue. The data are presented as the means and SDs (n = 3). Scale bar: 20 μm. Statistical significance was determined by two-tailed Student’s t-test. ***P < 0.005. c. Representative images (upper) and statistical quantification (lower) of apoptotic and necrotic cell populations in ESCC cell lines, as determined by Annexin-V FITC/PI staining and flow cytometry. Cells with a FITC⁻ and PI⁻ signature were considered viable. Cells with a FITC⁺ and PI⁻ or a FITC⁺ and PI⁺ signature were considered nonviable. The data are presented as the means and SDs (n = 3). Statistical significance was determined by two-tailed Student’s t-test. **P < 0.01. d. Representative images (upper) and statistical quantification (lower) of ROS production assay results in ESCC cells. The indicated cells were stained with carboxy-H₂DCFDA and observed under a fluorescence microscope. H₂DCFDA: green, Hoechst 33342: blue. Scale bar: 20 μm. The data are presented as the means and SDs (n = 3). Statistical significance was determined by two-tailed Student’s t-test. **P < 0.01. e-f. Representative images (left) and statistical quantification (right) of ROS production assay results in Eca109 (e) and TE-1 (f). The indicated cells were preincubated with different ROS inhibitors, stained with carboxy-H₂DCFDA and observed under a fluorescence microscope. H₂DCFDA: green, Hoechst: blue. Scale bar: 20 μm. The data are presented as the means and SDs (n = 3). Statistical significance was determined by one-way ANOVA. **P < 0.01