Fig. 6.

Insulin-induced activation of Akt/mTOR/p70S6K and ERK signaling pathways are more pronounced in PaSC cultured in 25 mM glucose compared with 5 mM glucose. imPaSCs were cultured in DMEM media containing 5 mM glucose and 10% FBS for 2 days, starved in serum-free medium for 4 h, and then cultured in 5 mM glucose- or 25 mM glucose-containing media with or without 100 nM insulin for various periods of time (1–24 h). A: cell lysates were analyzed for the indicated targets, and representative immunoblots are shown in A. β-Actin was used as loading control. B: graphs show protein levels of phosphorylated IGF-1R/IR, p70S6K, Akt, and ERK determined by densitometry. As indicated in the graphs, insulin-induced Akt and ERK phosphorylation was more pronounced in cells cultured in media containing 25 mM glucose compared with media containing 5 mM glucose. Data in graphs represent means ± SE from 3 independent experiments.