Fig 1: Fatty acid metabolism is essential for efficient SARS-CoV-2 replicationFASN CRISPR KO Caco2 cells and corresponding NT Caco2 cells were pre-seeded in 96-well plates, grown to confluency, and then infected at an MOI 0.01 in minimal media. Post-adsorption, cells were maintained in either 2% FBS DMEM (A) or 1% fatty-acid free (FAF)-BSA DMEM (B). Supernatants were collected at 1, 24, 48, 72, and 96 hpi and viral titers were determined by plaque assay. Protein samples were obtained from cell monolayers and analyzed by western blot to confirm FASN knockout and look for changes in SARS-CoV-2 N levels. FASN KO and WT cells were pre-seeded and infected as previously described. Post adsorption, inoculum was removed and replaced with 2% FAF-BSA DMEM or 2% FAF-BSA DMEM containing 250 μM palmitic acid + 250 μM oleic acid (C). Supernatants were collected at 1, 24, 48, and 72 hpi and viral titers were determined via plaque assay. The data represent the mean and SE of three replicates.