Y294002

Deferoxamine (DFO), CoCl₂, 2-methoxyestradiol (2-ME), N-acetyl-L-cysteine (NAC), MitoTEMPO, U0126, 5-aza-2’-deoxycytidine (5-aza-dC), Y294002, SB203580, Ro31-8220, rapamycin, BML-275, trichostatin A (TSA) and 2’,7’-dichlorofluorescein diacetate (DCF-DA) were obtained from Sigma-Aldrich (St Louis, MO. USA), and MitoPQ, MitoSOX-Red and 2‐(N‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)amino)‐2‐deoxyglucose (2‐NBD‐glucose) were from Abcam (Cambridge, UK), Thermo Fisher Scientific (Waltham, MA, USA) and Life Technologies (Carlsbad, CA, USA), respectively. AR-C155858 and SP600125 were supplied by Tocris Bioscience (Bristol, UK), and, syrosingopine was obtained from Extrasynthese SA (Lyon, France), respectively. TX-402 was a gift from Dr. H. Nagasawa, Gifu Pharmaceutical University. rapamycin was purchased from LC Laboratories (Woburn, MA, USA); BAY11-7082 was from AdipoGen Life Sciences (San Diego, CA, USA); PF-4708671 and KU-0063794 were from Selleck Chemicals LLC (Houston, TX, USA), and chaetocin was from Enzo Life Science (Farmingdale, NY, USA).

EPCs were obtained from mononuclear cells (MNCs) as the previous reports [23 (link)]. MNCs were discontinued into -complement medium (Gibco, USA) with endothelial growth factors (Sigma-Aldrich, USA), 10% FBS (Gibco, USA) and seeded on fibronectin (50 μg/mL, Sigma-Aldrich, USA)-pre-coated 6-well dishes (Sigma-Aldrich, USA). Cells were cultured at the condition of 5% CO₂ and 37° C. Dead cells were removed after three days, and then the culture mediums were refreshed every three days. And the EPCs were identified and characterized by DiI-acLDL and bind fluorescein iso-thiocyanate UEA-1 using immunofluorescent analysis after two weeks. The SAG, cyclopamine, and Y294002 were purchased (Sigma-Aldrich, USA).