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Experimental validation in <t>primary</t> <t>HASMCs</t> with siRNA knockdown and osmotic stress controls (A) Confirmation of AMPK knockdown in primary HASMCs. Representative western blots and AMPK/GAPDH ratios in scrambled siRNA- and AMPK siRNA-transfected cells are shown (n = 6; * P < 0.05 vs. control). (B) Effects of Gin A (10 μM) and AMPK knockdown on AMPK phosphorylation in HASMCs exposed to HG (25 mM) for 24 h. Representative blots and p-AMPK/GAPDH ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. si-AMPK alone; & P < 0.05 vs. si-AMPK + Gin A). (C) Effects of Gin A and AMPK knockdown on HG-induced HASMC proliferation. Cells were exposed to normal glucose or HG (25 mM) with or without Gin A (10 μM) and with scrambled or AMPK-targeting siRNA, and proliferation was measured by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A with scrambled siRNA). (D) Osmotic control experiments in HASMCs. Cells were cultured for 24 h in normal glucose (5.5 mM), HG (25 mM), L-glucose (25 mM) or D-mannitol (25 mM), and proliferation was assessed by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control).
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Experimental validation in primary HASMCs with siRNA knockdown and osmotic stress controls (A) Confirmation of AMPK knockdown in primary HASMCs. Representative western blots and AMPK/GAPDH ratios in scrambled siRNA- and AMPK siRNA-transfected cells are shown (n = 6; * P < 0.05 vs. control). (B) Effects of Gin A (10 μM) and AMPK knockdown on AMPK phosphorylation in HASMCs exposed to HG (25 mM) for 24 h. Representative blots and p-AMPK/GAPDH ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. si-AMPK alone; & P < 0.05 vs. si-AMPK + Gin A). (C) Effects of Gin A and AMPK knockdown on HG-induced HASMC proliferation. Cells were exposed to normal glucose or HG (25 mM) with or without Gin A (10 μM) and with scrambled or AMPK-targeting siRNA, and proliferation was measured by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A with scrambled siRNA). (D) Osmotic control experiments in HASMCs. Cells were cultured for 24 h in normal glucose (5.5 mM), HG (25 mM), L-glucose (25 mM) or D-mannitol (25 mM), and proliferation was assessed by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control).

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Experimental validation in primary HASMCs with siRNA knockdown and osmotic stress controls (A) Confirmation of AMPK knockdown in primary HASMCs. Representative western blots and AMPK/GAPDH ratios in scrambled siRNA- and AMPK siRNA-transfected cells are shown (n = 6; * P < 0.05 vs. control). (B) Effects of Gin A (10 μM) and AMPK knockdown on AMPK phosphorylation in HASMCs exposed to HG (25 mM) for 24 h. Representative blots and p-AMPK/GAPDH ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. si-AMPK alone; & P < 0.05 vs. si-AMPK + Gin A). (C) Effects of Gin A and AMPK knockdown on HG-induced HASMC proliferation. Cells were exposed to normal glucose or HG (25 mM) with or without Gin A (10 μM) and with scrambled or AMPK-targeting siRNA, and proliferation was measured by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A with scrambled siRNA). (D) Osmotic control experiments in HASMCs. Cells were cultured for 24 h in normal glucose (5.5 mM), HG (25 mM), L-glucose (25 mM) or D-mannitol (25 mM), and proliferation was assessed by MTT assay (n = 6; * P < 0.05 vs. normal-glucose control).

Article Snippet: Primary HASMCs were purchased from Procell (Wuhan, China) and used between passages 2 and 4.

Techniques: Biomarker Discovery, Knockdown, Western Blot, Transfection, Control, Phospho-proteomics, MTT Assay, Cell Culture