Figure 1. Metformin inhibits PDGF-induced phenotypic switching and increases the expressions of lncRNA-ANRIL in cultured VSMCs. Cultured VSMCs were treated with PDGF (5 ng/ml) for 48 hours in presence or absence of metformin (1 mM). (A) The morphology of contractile phenotype in cells was determined by immunofluorescence analysis of alpha SMA (α-SMA). (B) Total cell lysates were subjected to perform western blot analysis of phosphorylated AMPK (pAMPK), total AMPK protein levels, α-SMA, KLF4, and myocardin. (C) The mRNA levels of the phenotypic switching markers, including calponin, smoothelin, osteopontin, collagen I, and collagen III were measured by real-time PCR. (D) The lncRNA-ANRIL level was assessed by real-time PCR. (E) AMPK activity was assayed by P32-ATP method. N is 5 in each group. *P<0.05 vs. Vehicle. #P<0.05 vs. PDGF.