Fig. 6

CircAAGAB increases radiosensitivity in MDA-MB-231 cells. A Representative diagram of flow cytometry in MDA-MB-231 cells overexpressing circAAGAB and treated with ionizing radiation (IR). Cells were transfected with circAAGAB plasmids and exposed to 10 Gy IR. Cells were then collected at 24 h after IR, fixed with 75% ethanol overnight, and stained with PI. B Quantification of cell cycle and apoptotic cells (sub-G1). C Representative diagram of flow cytometry with annexin-V and PI staining. Cells were transfected with circAAGAB plasmids and exposed to 10 Gy IR. Cells were then collected at 24 h after IR. D Quantification of early and late stages of apoptosis in MDA-MB-231 cells. E, G Western blotting of γH2AX (E) and caspase-3 (CASP3) (G) in MDA-MB-231 cells overexpressing circAAGAB and treated with IR. Cells were transfected with circAAGAB plasmids and exposed to 10 Gy IR. Cells were collected at 4 h after IR for γH2AX, and 6 h for CASP3. Loading controls: ACTB or GAPDH. F, H Quantification of γH2AX (F) and CASP3 (H) in MDA-MB-231 cells overexpressing circAAGAB. *P < 0.05, **P < 0.01, ***P < 0.001