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. 2022 Nov 14;13:6754. doi: 10.1038/s41467-022-34095-x

Fig. 9. Effects of chronic hyperglycaemia, koningic acid and PS10 on PDH signalling.

Fig. 9

a Representative Western blot of lysates from control (C) and diabetic islets (Db) cultured in the presence or absence of 10 µM S6-kinase inhibitor PF-4708671 (S6Ki) and then stimulated with 2 mM or 20 mM glucose for 1 h. Phosphorylated (p) PDHe1α. Loading control, α-tubulin. Uncropped blots in source data. b Quantitative densitometry analysis of p-PDHe1α/ α-tubulin (n = 3, 11 animals/genotype). Control islets (black), Diabetic islets (red), Control islets + S6Ki (blue), Diabetic islets + S6Ki (orange). Veh, vehicle (0.05% DMSO). c Representative Western blot of lysates from LG-cells, HG-cells and LG-cells cultured for 48 h with 5 µM koningic acid (KA), then stimulated with 2 mM or 20 mM glucose for 1 h. Phosphorylated (p) PDHe1α. Loading control, α-tubulin. Uncropped blots in source data. d Quantitative densitometry analysis of p-PDHe1α/α-tubulin (n = 4 biologically independent experiments). LG-cells (black), HG-cells (red), LG-cells+KA (blue). e Insulin secretion from HG-cells, and LG-cells cultured with or without 5 µM koningic acid (KA) for 48 h and subsequently stimulated acutely with 2 mM glucose (G) or 20 mM methyl pyruvate (Pyr) for 30 min (n = 3 biologically independent experiments). f Representative Western blot of lysates from control (C) and diabetic (Db) islets stimulated with 2 mM or 20 mM glucose ± 50 µM of the PDK inhibitor PS10 for 1 h. Uncropped blots in source data. g Insulin secretion from control and diabetic islets stimulated with 2 mM or 20 mM glucose (G), 20 mM glucose + 50 µM PS10, or 20 mM KCl for 1 h. Control islets (black, n = 4 animals), Diabetic islets (red, n = 4 animals). All panels show individual data points and 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.