Leupeptin

Boceprevir, Telaprevir, Narlaprevir, Grazoprevir, Asunaprevir, Simeprevir, Leupeptin, VBY-825, Balicatib, CA-074, Calpeptin, E-64, E-64c, JPM-OEt, LY-3000328, Odanacatib, CPI (Cysteine Protease Inhibitor), Aloxistatin, Cinanserin, MDL-28170, MG-101, ONO-5334, (±)Alliin, and Z-LVG-CHN2 were purchased from MedChemExpress, LLC. (www.medchemexpress.com). DiscoveryProbe™ FDA-approved Drug Library (23 plates of 96-well, 1971 compounds) were purchased from APExBIO (https://www.apexbt.com).

A7r5 cells (Rat thoracic artery smooth muscle cells) were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Item No: ZQ0139). Cells were cultured in DMEM (Hyclone, United States) containing 10% fetal bovine serum (FBS) (Gibco, United States) and 1% penicillin/streptomycin (Gibco, United States). Cells were incubated under 5% CO₂ at 37°C in humidified incubators. To induce dedifferentiation of VSMCs in vitro, A7r5 cells were starved with serum-free medium for 48 h, and were then stimulated with 10% FBS serum as previously described (Zhang et al., 2020b (link)). The cells were harvested at 0, 24, 48, and 72 h for subsequent detection. The A7r5 cells were infected with adenovirus expressing ERα (Ad-ERα), adenovirus expressing OMI (Ad-OMI) or the negative control adenovirus (Ad-NC) in normal culture medium for 48 h. After 48 h or 72 h of siRNA/virus treatment, RNA or protein was extracted for further assays. The siRNA against OMI and the negative control siRNA transfection was performed using Lipofectamine RNA interference (RNAi) max reagent (Thermo Fisher Scientific, United States) according to the manufacturer’s protocol. For inhibitor treatment, 3-MA (10 mM), FCCP (5 μM), MG132 (5 μM) and leupeptin (50 μM) were purchased from MedChemExpress Co., Ltd. (New Jersey, United States).