Fig. 4

Overexpression of FTO attenuated cisplatin-induced granulosa cell apoptosis and promoted its proliferation. A The transfection efficiency of FTO was measured by qRT-PCR. B-C The mRNA expression of Bcl-2 and BAX in transfected granulosa cells was assessed using qRT-PCR and normalized to GAPDH. D-G The protein expression of FTO, BAX, and Bcl-2 was determined by western blotting with β-actin as a loading control in different groups. (H-K) Western blotting results showed that overexpression of FTO in injured granulosa cells could restore the cisplatin-induced change of FTO, Bcl-2, and BAX. L-M EdU labeling and cck-8 assay indicated that cells viability was inhibited in cisplatin-induced injured cells. However, transfection of FTO in injured cells, the viability was increased. N-O Flow cytometry assays were performed to determine the apoptotic rate in different groups. The results showed that cell apoptotic rate was increased in the cisplatin-induced injured cell, while co-transfection of FTO could restore it. One representative experiment of three independent experiments is shown. The results represent the mean ± SEM from three independent experiments. Student’s t-test and one-way ANOVA were applied to compare the two experimental and multiple groups, respectively. (*P < 0.05, **P < 0.01, ***P < 0.001, ns, no significance)