Figure 2.

Reduced m6A methylation-mediated by ALKBH5 suppresses osteosarcoma cell growth and promotes cell apoptosis. a. ALKBH5 overexpression was confirmed by western blot in U2OS and KHOS cell lines. b. ALKBH5 up-regulation reduced m6A mRNA methylation. c. Cell viability was assessed by CKK-8 assay. d. The percentage of apoptosis was determined by FCM analysis. e. TUNEL staining was detected from osteosarcoma cells with transfection of Lv-ALKBH5, positive cells were labeled with TUNEL (green) (magnification × 400) . f. Cell cycle analysis was performed using FCM, which revealed osteosarcoma cells were mostly arrested in the G0/G1 phase. g. SiRNA was used to down-regulate ALKBH5, and ALKBH5 down-regulation was confirmed by western blot. h,i. Flow cytometry (h)and TUNEL (i) assay were carried out to confirm the reduction of apoptosis. j. Cell cycle assay showed that there was an increase in the number of dividing tumor cells following the knockdown of ALKBH5. The data represent the mean± S.D. of three independent experiments. ANOVA or t-tests was used for statistical analysis. *P < 0.05 or **P < 0.01 indicates a significant difference between the indicated groups.