Figure 3
Sirt1 downregulation by miR-204 promotes ER stress. (A) Immunoblots for ER stress markers in HUVECs transfected with siRNA Sirt1 (20 pmol) or treated with Tunica. Control cells were transfected with a scrambled siRNA (siRNA SC) or treated with vehicle (Veh). Data is representative of three independent experiments. (B) Immunoblots for ER stress markers and Sirt1 in HUVECs transfected with Sirt1 adenovirus (Ad Sirt1) and treated with Tunica. Control cells were infected with an adenovirus expressing LacZ (Ad LacZ) and treated with Veh. Data is representative of three independent experiments. (C) Immunoblots for ER stress markers in HUVECs transfected with miR-204 mimic (50 nM for 48 h) and infected with Ad Sirt1. Data is representative of three independent experiments. Sirt1 quantified by qPCR in thoracic aorta (D) and by qPCR and immunofluorescence in MRAs (E,F) of mice systemically infused with miR-204 I (0.2 mg/mouse/week) and treated with Tunica. Sirt1 mRNA is normalized to GAPDH and expressed relative to miR SC + Veh. *p < 0.05 vs miR SC + Veh, miR-204 I + Tunica, and miR-204 I + Veh. #p < 0.05 vs miR SC + Tunica. vWF: Von Willebrand Factor.
