The regulation of PHB by gonadotropin. (A) Total RNAs of undifferentiated and differentiated granulosa cells were extracted and mRNA abundance of PHB was analyzed by PCR. p450scc and aromatase mRNA levels were assessed as differentiated marker and Actin was used as loading control. (B) Undifferentiated and differentiated granulosa cells were cultured with FSH (100 ng/ml) for designated time period and the contents of PHB, pAkt and total Akt were examined by Western blot. (C) Undifferentiated and differentiated granulosa cells were cultured with FSH (0–200 ng/ml) for 24 h. PHB, pAkt and total Akt contents were examined by Western blot. Representative immunoblots are shown and data are presented as mean ± SEM of three independent experiments. B-C, one-way ANOVA, followed by Bonferroni test. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (compared with FSH = 0 or FSH at 0 h).