Total proteins were extracted as described previously [
12]. Cells were lysed in radioimmunoprecipitation assay buffer (Beyotime; Haimen, China), and protein concentrations were measured through bicinchoninic protein assay kit (Pierce, Appleton, WI, USA). Equal amounts of protein were separated through SDS-PAGE and subsequently transferred to a PVDF membrane (Millipore; Burlington, MA, United States). After the membrane was blocked for 1 h using 5% skim milk, it was incubated with primary antibody overnight at 4 °C. Finally, the membrane was incubated with respective secondary antibodies (Santa Cruz, United States) for 1 h. Signals were detected using enhanced chemiluminescence reagents (Pierce; Waltham, MA, United States). Sources of antibodies and concentrations used were as follows: rabbit anti-PKM2 (1:1000, CST, USA), rabbit anti-phospho-ATM (Ser1981) (1:1000, CST, USA), rabbit anti-phospho-BRCA1 (Ser1524) (1:1000, CST, USA), rabbit anti-phospho-Chk1 (Ser345) (1:1000, CST, USA), rabbit anti-phospho-Chk2 (Thr68) (1:1000, CST, USA), rabbit anti-phospho-histone H2AX (Ser139) (1:1000, CST, USA), mouse anti-cycline B1 (1:1000, Santa Cruz, USA), mouse anti-phospho-p53 (Ser15) (1:1000, CST, USA), rabbit anti-cleaved caspase 3 (1:1000, CST, USA), rabbit anti-caspase 3 (1:1000, CST, USA), rabbit anti-cleaved caspase 9 (1:1000, CST, USA), mouse anti-caspase 9 (1:1000, CST, USA), rabbit anti-Bcl2 (1:1000, CST, USA), rabbit anti-Bax (1:1000, CST, USA), rabbit anti-NANOG (1:1000, Abcam, USA), rabbit anti-OCT4 (1:1000, CST, USA), rabbit anti-SOX2 (1:1000, CST, USA), rabbit anti-KLF4 (1:1000, CST, USA), mouse anti-ABCG2 (1:1000, CST, USA), mouse anti-Bmi1 (1:1000, Santa Cruz, USA), rabbit anti-GAPDH (1:1000, CST, USA). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Quantity One Software (Bio-Rad) was used to analyze the intensity of blots.