Figure 5. The effect of insulin in S6K Thr³⁸⁹ phosphorylation with or without pre-exposure of low and high dose testosterone.

(A). The effect of insulin alone on S6K Thr³⁸⁹ phosphorylation after insulin treatment for 10, 20, 30, 60, and 120 minutes; Insulin significantly increased S6K Thr³⁸⁹ phosphorylation after 60 minutes; (B). Cells were pre-exposed to low-dose (20ng/ml) or high-dose (200ng/ml) of testosterone for 16 hours before insulin treatment. Compared to non-testosterone treated cells, S6K Thr³⁸⁹ phosphorylation was significantly increased with low-dose testosterone treatment. Representative western blots are shown above the bar graphs. The four experimental groups are: control cells without insulin (C−I), control cells plus insulin (C+I), testosterone exposed cells without insulin (T−I), and testosterone exposed cells plus insulin (T+I). The phosphorylated signal of S6K Thr³⁸⁹ is normalized to total S6K in the sample.