Figure 4. NR4A2 knockdown inhibited H₂O₂-induced autophagy and apoptosis in CSCs.

CSCs were challenged with indicated concentrations of H₂O₂ for 5 h. (A) Transcription of NR4A2 was observed by qPCR. (B) Representative immunofluorescent images showed the expression of NR4A2 in CSCs treated with H₂O₂. Scale bar=20 μm. (C) NR4A2 protein levels were determined by western blot. (D and E) CSCs were transfected with NR4A2 siRNA for 48 h, then the mRNA and protein were extracted for qPCR detection (D) and western blot (E). (F) CSCs were transfected with NR4A2 siRNA for 43 h followed by H₂O₂ treatment for another 5 h. cleavage of caspase 3 and PARP1, LC3-II and P62 were detected by western blot. neg, negative control, siN, siNR4A2, ctr, control. *P < 0.05; **P < 0.01; n=3.