Cells Lines

Multiple cell lines were acquired in this study to determine permissiveness to SARS-CoV-2 for use in drug screens. Vero-E6 cells were acquired from the American Type Culture Collection (ATCC) and maintained in high glucose DMEM (Gibco, Waltham, MA) supplemented with 10% FBS (R&D Systems, Minneapolis, MN), 1X GlutaMAX (Gibco), and 1X PenStrep (Gibco) [D10 media]. Huh-7 and HPMEC cells were obtained from Dr. Eva Harris (UC Berkeley) and maintained in D10 media or endothelial growth medium 2 (EGM-2) using the EGM-2 bullet kit from Lonza, respectively. LNCaP, HaCaT, Caco-2, Calu-3, HCT-116, and A549 cells were obtained from the UC Berkeley Cell Culture Facility and maintained in D10 media. NCI-H1437 and RD cells were obtained from the Cell and Genome Engineering Core at UCSF via Dr. Olga Gulyaeva (UCSF) and Dr. Michael T. McManus (UCSF) and maintained in D10 media. HBEC-30KT and BEAS-2B cells were obtained from Dr. Neil Tay (UCSF) and Dr. Michael T. McManus (UCSF) via Dr. Patrick Mitchell (UC Berkeley) and Dr. Russell Vance (UC Berkeley) and maintained in defined keratinocyte serum free medium (catalog #10744019, ThermoFisher Scientific) and Advanced RPMI containing 5% FBS, 1% l-glutamine, and 1X PenStrep, respectively. Huh-7.5.1 cells were obtained from Dr. Andreas Puschnik (Chan Zuckerberg Biohub) and maintained in D10 media. All cells were maintained in a CO₂ incubator at 37 °C with 5% CO₂. HPMEC/hACE2 and A549/hACE2 cell lines were produced by transducing parental cells with lentivirus encoding the human ACE2 gene followed by puromycin selection (2 μg/mL) for three passages. The hACE2 encoding plasmid was a gift from Hyeryun Choe (Addgene plasmid # 1786; http://n2t.net/addgene:1786; RRID:Addgene_1786).⁵⁵