2 apb

Neonatal Sprague-Dawley rats (1 to 3 days old) were exposed to hypothermia until they became unresponsive and were then euthanized by cervical dislocation. Primary culture of NRCMs was performed according to previously published method 24 (link). The cells were seeded at a density of 5 x 10⁶ cells/ml in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 0.1 mM 5-bromodeoxyuridine. After 24 h, NRCMs were subjected to various treatments as described below. To assess the effects of EndoA2 on cardiac injury, the cells were transfected with EndoA2 siRNA or Ad-EndoA2. After transfection for 36 h, the cells were stimulated with oxygen and glucose deprivation (OGD) for another 12 h. Incubation of sodium phenylbutyrate (4-PBA, Sigma, Boston, MA), Cyclosporin A (CsA, Selleck, Houston, TX), 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA, Selleck, Houston, TX) or 2-Aminoethoxydiphenylborate (2-APB, Selleck, Houston, TX) was 2 h before OGD stimulation. Cardiomyocytes were cultured in serum-free, glucose and sodium pyruvate- free DMEM (Sigma, USA) at 37 °C in an anoxia chamber (InVivo 500, Skinn Life Science) saturated with 94% N₂/5% CO₂/ 1% O₂.

Vero E6 cell line was obtained from American Type Culture Collection and maintained in minimum Eagle’s medium (Gibco Invitrogen) supplemented with 10% fetal bovine serum (FBS; Gibco Invitrogen) and 1% antibiotic/antimycotic (Gibco Invitrogen), at 37 °C in a humidified 5% CO₂ incubator. Huh7 cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco Invitrogen) supplemented with 10% FBS and 1% antibiotic/antimycotic (Gibco Invitrogen), at 37 °C in a humidified 5% CO₂ incubator.
SARS-CoV-2 (nCoV-2019BetaCoV/Wuhan/WIV04/2019) was propagated in Vero E6 cells^{2 (link)}, and viral titer was determined by 50% tissue culture infective dose (TCID50) as described in our previous study^{5 (link)}. All the infection experiments were performed in a biosafety level-3 (BSL-3) laboratory.
Benidipine HCI (Selleck Chemicals, no. S2017), amlodipine besylate (Selleck Chemicals, S1813), cilnidipine (Selleck Chemicals, S1293), nicardipine HCl (Selleck Chemicals, S4181), nifedipine (Selleck Chemicals, S1808), isradipine (Selleck Chemicals, S1662), nimodipine (Selleck Chemicals, S1747), nisoldipine (Selleck Chemicals, S1748), felodipine (Selleck Chemicals, S1885), 2APB (Selleck Chemicals, S6657), BAPTA-AM (Selleck Chemicals, S7534), and CQ (Sigma-Aldrich, no.C6628) were purchased from indicated companies.