As described in previous studies, total protein was extracted from the NRVMs and heart tissue 21. Briefly, proteins were separated by electrophoresis on an SDS-polyacrylamide gel and transferred wet to a nitrocellulose filter. Membranes were incubated with anti-FoxO4 (1:100, sc-373877, mouse monoclonal, Santa Cruz, USA) antibody overnight at 4°C with the following secondary antibodies for 1 h at room temperature in the dark. The images were captured by the Odyssey CLx Infrared Imaging System (LI-COR Biosciences, Lincoln, NE, USA). Anti-β-actin (1:1000, sc-8432, mouse polyclonal, Santa Cruz, CA, USA) antibody is internal control.

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