Fig. 4.

mTOR siRNA inhibits hCG-induced steroidogenic enzyme expression. T-I cells were transfected with 100 nm of control siRNA or mTOR siRNA for 48 h and then treated without or with hCG for another 24 h. A, Proteins (30 μg) were separated by SDS-PAGE (4–20%) and then immunoblotted with mTOR, StAR, P450scc, HSD3B1, and P450c17 antibodies. Protein loading was monitored by stripping and reprobing the same blot with antibody for GAPDH. The graphs (B–F) represent densitrometric scans of mTOR, StAR, P450scc, HSD3B1, and P450c17 protein expression normalized for GAPDH as seen in A. The blots in A represent one of three separate experiments, and the graphs represent the mean of three experiments. Error bars represent mean + se. ***, P < 0.001 vs. control siRNA. a, and c represent significant differences (P < 0.05 and P < 0.001, respectively), compared with control siRNA plus hCG treatment.