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. Author manuscript; available in PMC: 2014 Jan 4.
Published in final edited form as: Biochem Biophys Res Commun. 2012 Nov 15;430(1):352–357. doi: 10.1016/j.bbrc.2012.11.010

Fig. 2. Enhanced metformin-induced mTORC1 inhibition in PDAC cells exposed to physiological glucose concentrations.

Fig. 2

A) Cultures of MiaPaca-2 and PANC-1 cells were incubated without (−) or with 1 mM metformin for 16 h in DMEM containing 5 mM or 25 mM glucose, as indicated. Then, the cells were stimulated for 2 h with 5 nM neurotensin (NT), 10 ng/ml insulin (Ins) or their combination (NT+Ins), lysates were analyzed by immunoblotting with the following phospho antibodies: S6K Thr389 (pS6K), S6 Ser235/236 (pS6) and ACC Ser79 (pACC). Total ACC was used to verify equal gel loading. Similar results were obtained in 3 independent experiments. B). Mia PaCa-2 cells were incubated with DMEM containing 5 mM glucose either in absence or presence of 0.05mM or 0.1 mM metformin for 16 h. Then, the cells were treated with NT+Ins, as in panel A, and lysates were analyzed by immunoblotting. Similar results were obtained in 6 independent experiments. Results are expressed as the percentage of maximum mean ±SEM, n=6 (S6K) or as the fold increase over basal as indicated. P values were determined using the t-test (Sigma Plot 12.) *p<0.05; **p<0.001; n=6.