Primary antibodies against sirt1

Sirtinol was purchased from Selleck Chemicals LLC (Houston, TX, USA). Culture medium and its supplements including antibiotics and fetal bovine serum (FBS) were purchased from Euroclone (Italy). Primary antibodies against SIRT1 (Abcam, Cambridge, UK), HIF-1α (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and tubulin (Sigma-Aldrich, Milan, Italy) were used. Sirtinol was dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich) to the appropriate concentrations according to reported procedures. DMSO was also present in the corresponding control.

BAK (purity ≥ 98%) was purchased from Winherb Medical Co. (Shanghai, China). Streptozotocin (STZ) and EX527 were purchased from Sigma-Aldrich (St. Louis, MO, USA). The In Situ Cell Death Detection Kit was purchased from Roche Biochemicals (Mannheim, Germany) to detect the cell apoptosis. Kits for determining glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) activities, and malondialdehyde (MDA) content were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) was purchased from Beyotime Institute of Biotechnology (Shanghai, China) to detect intracellular ROS generation. Dihydroethidium (DHE) was purchased from Invitrogen (Carlsbad, CA, USA) for detecting ROS generation in the myocardium. Primary antibodies against p-Smad3, t-Smad3, and β-actin were purchased from Cell Signaling Technology (Boston, MA, USA). Primary antibodies against SIRT1, Nrf2, transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), and Histone H3 were purchased from Abcam (Cambridge, MA, USA). The rabbit anti-goat and goat anti-mouse secondary antibodies were obtained from Zhongshan Company (Beijing, China).