Western blotting

The GCs and rat ovarian tissue were lysed in RIPA lysis buffer (Shenggong, Shanghai, China) containing a protease inhibitor cocktail (Roche, Basel, Switzerland) on ice, after which the supernatant was collected. After being quantified using the Bradford assay (Beyotime), 30 μg of proteins were loaded onto 10–12.5% sodium dodecyl sulfate polyacrylamide gels. After being transferred to nitrocellulose membranes and blocked with 5% non-fat milk for 1 h at room temperature, membranes were incubated with primary antibody and secondary antibody. The peroxidase activity of bands was detected using the chemiluminescent detection kit (Millipore Sigma, Burlington, MA, USA). The primary and secondary antibodies used were: NCOA4 (raised in rabbit, 1:500; Abcam), FTH1 (raised in rabbit, 1:1000; Abcam), GPX4 (raised in rabbit, 1:1000; Abcam), and GAPDH (raised in mouse, 1:10000; Proteintech, Wuhan, China).