Fig. 5.

Examination of RagA overproduction in the activation of mTOR-p70S6K pathway in LPS-treated PC12 cells. a LPS induced the activation of RagA, mTOR, and p70S6K. After 24 h drug treatment, PC12 cells were lysed and analyzed by western blotting with antibodies against RagA, mTOR, phospho-mTOR, p70S6K, and phospho-p70S6K, whereas GAPDH was analyzed as control. Representative blot was shown. b Quantitative analysis of protein expression. Western blots in a were determined by a densitometric method. The signals (means ± SD) from three independent experiments were analyzed by one-way ANOVA, followed by post hoc Tukey’s test. *p < 0.05; **p < 0.01; ***p < 0.001 (LPS vs control). c Western blotting analysis for siRNA-mediated RagA silencing. PC12 cells were transfected with RagA-specific and negative control siRNAs for 72 h. The transfected cells were treated with 1 μg/ml of LPS for 24 h. The cells were analyzed by western blotting for RagA expression, whereas GAPDH was analyzed as control. Representative blot was shown. d Quantitative analysis of RagA silencing efficiency. Western blots in c were determined by a densitometric method. The signals (means ± SD) from three independent experiments were analyzed by one-way ANOVA, followed by post hoc Tukey’s test. *p < 0.05; **p < 0.01; ***p < 0.001 (LPS vs control/ LPS + RagA siRNA/ LPS + negative siRNA)