Figure 4.

Inhibitory effects of rapamycin on insulin-induced mTOR activity, CRMP-2 and synapsin-I expression and neuronal growth in HT-22 cells. Cells were treated with insulin (10 nM) in the absence and presence of rapamycin (5 μM) for 24 and 48 h. The p-S6K/S6K, p-S6/S6, CRMP-2 and synapsin-I levels were analyzed by Western blotting, and subsequently quantified (A–E). Viable cells were assayed using a CCK-8 (E). Representative data for microscopic observation (200×, Scale bar, 100 μm) of cells are presented (F). The quantitative data are the mean ± SEM of six experiments. Significance values indicate *p < 0.05, and **p < 0.01 vs. vehicle control: ^†p < 0.05 and ^††p < 0.01 vs. insulin control.