Figure 5. Attenuation of VPS34 kinase activity and fatty acid metabolism inhibit SARS-CoV-2 replication in human airway epithelial cell line.

Calu-3 cells were plated onto a 96-well plate and allowed to reach 95% confluency. Cells were then pre-treated with a range of concentrations of A-B) VPS34-IN1, C-D) PIK-III, E-F) Orlistat, G-H), Triacsin C, DMSO, or mock-treated with media alone for 1 hour then infected with SARS-CoV-2 at an MOI of 0.01. Supernatants were collected at 48 h.p.i. and virus was quantified by plaque assay on VeroE6 cells. The data is reported as plaque forming units per milliliter (pfu/ml) (left panels). Cell viability over 48 hours was determined in parallel. Percent inhibition, IC50, and IC90 were calculated from the plaque assay data and plotted with the cell viability data (right panels). The dotted line labeled DMSO indicates the level of virus growth in the DMSO control. The dotted line labeled LOD indicates the limit of detection of the plaque assay.