SARS-CoV-2 Propagation and Drug Screening

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.