Telmisartan Modulates Cardiac Signaling

Unfixed frozen hearts from WT mice treated with telmisartan for two weeks were homogenized in lysis buffer and centrifuged^{16 (link)}. Following protein quantification, samples were resolved using SDS-PAGE and transferred onto nitrocellulose membrane. The membrane was cut just below 100kDs and the top and bottom sections were blocked and probed overnight with primary antibodies for eNOS (1:1000, Cell Signaling, Cat# 32027) on the top membrane, and T-AKT (1:1000, Cell Signaling, Cat# 2920), p-AKT (1:1000, Cell Signaling, Cat# 4060), Cav-1 (1:1000, Santa Cruz Biotechnology, Cat# sc-894) and GAPDH (1:4000, Cell Signaling, Cat# 2118) on the bottom membrane, followed by incubation with goat anti-rabbit AlexaFluor700 (1:5000, Thermo Fisher Scientific, Cat# A-21038) and goat anti-mouse AlexaFluor800 (1:5000, Thermo Fisher Scientific, Cat# A-32730) secondary antibodies before imaging with a Li-COR scanner. The Li-COR software was used to designated the area of interest to be scanned for the bottom membrane.