Figure 5. Adenovirus-mediated knockdown of lncRNA-ANRIL prevents the anti-atherosclerotic effects of metformin in Apoe−/− mice. (A) The protocol of animal experiments in Apoe−/− mice. Male Apoe−/− mice were injected with adenovirus containing scramble shRNA or lncRNA-ANRIL shRNA to silence lncRNA-ANRIL gene expression. Two weeks later after injection, mice received western diet plus metformin (150 mg/kg/day) in drinking water for 4 weeks. At the end of experiments, all mice were sacrificed under anesthesia. (B) The aortic root tissue was subjected to fluorescence in situ hybridization to detect lncRNA-ANRIL expression in green and the morphology analysis of aortic root by Oil Red staining and HE staining. (C) The expressional level of lncRNA-ANRIL and AMPK activity in aortic lesion tissue were assayed. (D) Quantitative analysis of atherosclerotic lesion size in aortic root by Oil Red staining and HE staining. N is 10-15 in each group. *P<0.05 vs. scramble shRNA alone. #P<0.05 vs. scramble shRNA plus metformin.