Research Paper Volume 12, Issue 9 pp 8067—8083

BMAL1 knockdown triggers different colon carcinoma cell fates by altering the delicate equilibrium between AKT/mTOR and P53/P21 pathways

Figure 7. HCT116-shBMAL1 cells temporarily increased apoptosis and P53 activation after lentivirus transduction. (A) Flow cytometry analysis with Annexin V-APC and propidium iodide staining were applied to determine apoptosis ratio in different shRNA (shBMAL1 or shScr) transduced cells 48h after lentivirus transduction. Upper panels, a representative distribution of three independent experiments is shown. Lower panels, Graphs represent the percentage of early apoptosis cells (Q4) and total apoptosis cells (Q2+Q4) (3 independent for each analysis). A significant increase of apoptosis ratio is detected only in HCT116 cells after shBMAL1 transduction (**p<0.01). (B) Flow cytometry analysis with P53-PE and Hoechst 33342 staining revealed that the nuclei of HCT116 shBMA1 transduced cells (GFP positive population) exhibited an increased P53 expression compared to the nuclei of HCT116 shScr transduced cells (n=3; **p<0.01). Right, Graphs represent the mean of P53 nuclei expression from three independent experiments. All data are shown as means ± SEM.