Fig. 5: Chronic hyperglycaemia decreases AMPK and increases mTORC1 signalling.

a Representative Western blot of lysates from control (C) and diabetic (Db) islets stimulated with 2 mM or 20 mM glucose for 1 h. Phosphorylated (p) and total AMPK, Raptor, S6 and 4E-BP1. Uncropped blots in Source data. b–e Quantitative densitometry analysis of p-AMPK/AMPK (b, n = 4, 12–15 animals/genotype), p-Raptor/Raptor (c, n = 5, 14–17 animals/genotype), p-S6/S6 (d, n = 4, 12–15 animals/genotype) and p-4E-BP1/4E-BP1 (e, n = 4, 12–15 animals/genotype). Control, black bars. Diabetic, red bars. f Representative Western blot of lysates from LG-cells and HG-cells stimulated with 2 mM or 20 mM glucose for 1 h. Phosphorylated (p) and total AMPK, Raptor, S6 and 4E-BP1. Uncropped blots in Source data. g–j Quantitative densitometry analysis of p-AMPK/AMPK (g, n = 5), p-Raptor/Raptor (h, n = 4 biologically independent experiments), p-S6/S6 (i, n = 3 biologically independent experiments) and p-4E-BP1/4E-BP1 (j, n = 4 biologically independent experiments). LG-cells, black bars. HG-cells, red bars. k Representative Western blot of lysates from LG-cells, HG-cells and LG-cells cultured for 48 h with 5 µM koningic acid (KA), and then stimulated with 2 or 20 mM glucose for 1 h. Phosphorylated (p) and total AMPK and S6. Uncropped blots in source data. l, m Quantitative densitometry analysis of p-AMPK/AMPK (l, n = 3 biologically independent experiments) and p-S6/S6 (m, n = 3 biologically independent experiments). LG-cells, black bars. HG-cells, red bars. LG-cells + KA, blue bars. All panels show individual data points plus mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed unpaired Student’s t test. Source data are provided as a Source Data file.