Human HEK293T (American Type Culture Collection [ATCC]), HeLa (NIH HIV Reagent Program, Division of AIDS, NIAID, NIH from Dr. Richard Axel), HEK293T-ACE2 (BEI resources), and VeroE6 (ATCC) cells were cultured in complete medium (Dulbecco’s Modified Eagle Medium [DMEM, ThermoFisher Scientific] supplemented with 10% fetal bovine serum [FBS, ThermoFisher Scientific], 1% Penicillin-Streptomycin [ThermoFisher Scientific] and 1% L-glutamine [ThermoFisher Scientific]). A549-ACE2 (BEI resources) cells were maintained in complete medium with 50 ng/mL blasticidin S HCl (ThermoFisher Scientific). HEK293T-ACE2 stably expressing pQCXIP-BST2 or pQCXIP, HEK293T-ACE2-BST2-ATG5KO and HEK293T-ACE2-BST2-LentiCRISPRv2 cells were grown in complete medium with 1 μg/mL of puromycin (ThermoFisher Scientific). A549-ACE2 cells stably expressing pQCXIP-BST2 or pQCXIP were maintained in complete medium with 50 ng/mL blasticidin S HCl and 1 μg/mL of puromycin. Viability of cells was measured after each transfection. Only cells with viabilities > 90% were considered for further experiments.
Shi Y., Simpson S., Chen Y., Aull H., Benjamin J, & Serra-Moreno R. (2024). Mutations accumulated in the Spike of SARS-CoV-2 Omicron allow for more efficient counteraction of the restriction factor BST2/Tetherin. PLOS Pathogens, 20(1), e1011912.
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