Figure 6

Effect of AMPKα1, AMPKα2, and SIRT1 siRNAs on anthocyanin-stimulated AMPK signalling in HGECs. Protein lysates (10 μg) from cultured HGECs were separated by SDS-PAGE and analysed by western blotting. Cultured HGECs were transfected with 40 nmol/L control siRNA or 40 nmol/L AMPKα1, AMPKα2, or SIRT1 siRNAs using a transfection reagent (Lipofectamine 2000). At approximately 24 h after transfection, the levels of phospho-AMPK Thr¹⁷², total AMPK, SIRT1, and PGC-1α signalling in low-glucose medium were analysed. a–d Representative western blots (a) and quantitative analyses of the results b–d. *p < 0.05 and **p < 0.01 compared with the other groups. Cultured HGECs were transfected with 50 nmol/L control siRNA or 50 nmol/L AMPKΑ1, AMPKΑ2, or SIRT1 siRNAs and stimulated with anthocyanin in high-glucose medium. At approximately 24 h after stimulation, the levels of phospho-AMPK Thr¹⁷², total AMPK, SIRT1, PPARα, PGC-1α, and ERR-1α signalling in high-glucose medium were analysed. e–j Representative western blots of phospho-AMPK Thr¹⁷², total AMPK, PPARα, PGC-1α, ERR-1α, and β-actin levels (e) and quantitative analyses of the results (f–j). *p < 0.05 and **p < 0.01 compared with high-glucose medium. HG high glucose.