Figure 7. Immunoblot for phospho-AMPK, total AMPK, and PGC-1α in AMPKα1 siRNA, AMPKα2 siRNA, or PGC-1α siRNA knock-down mesangial cells (NMS) in a high-glucose environment with fenofibrate treatment.

The cultured mesangial cells were transfected with 50/l control siRNA or 50 nmol/l Ampkα1, Ampkα2, or PGC-1α siRNA using transfection reagent (G-Fectin) and stimulated with fenofibrate in high-glucose media. Approximately 48 hr after transfection, the levels of phospho-Thr¹⁷² AMPK, total AMPK, SIRT1, and PGC-1α signaling in the high-glucose media stimulated with fenofibrate, were analyzed. Protein lysates (10 µg) from the cultured mesangial cells were separated by SDS-PAGE and analyzed by Western blotting. A–C) Representative Western blot analysis of phospho-Thr¹⁷² AMPK, total AMPK, PGC-1α, and β-actin levels, (A) and the quantitative analyses of the results are also shown: phospho-Thr¹⁷² AMPK and total AMPK (B); PGC-1α (C). *p<0.05 compared with other groups. Abbreviations: HG, high glucose; Feno, fenofibrate; pAMPK, phospho-Thr¹⁷² AMPK.