Extended Data Fig. 4: Confirmation of gene editing efficiency for LOF cells and loss of AP1 adaptor complex components decreases Spike glycoprotein-pseudotyped viral entry.
a. Next generation sequencing was performed to analyze indel percentage in the indicated LOF cell Calu-3 lines. Graph depicts the indel percentage for each guide compared to NTG editing efficiency at the same locus. b. Western blot analysis for a representative KO target ROCK1 indicates successful depletion of the target gene for two guides. One western blot was conducted in this experiment, then re-probed for β-actin as a loading control c. Time-course of VSV-CoV-2-S infection of NTG, AP1G1, AP1B1 and AP1M2 KO cell lines. NTG, non-targeting guide. VSV-CoV-2-S, SARS-CoV-2 Spike-pseudotyped vesicular stomatitis virus encoding GFP. n = 3 biological replicates. d. Number of GFP + cells at 6 hpi, from the time-course shown in (c). Statistical significance was measured by individual two-sided t-tests comparing GOF lines to NTG lines, n = 3 biological replicates. e. Time-course of VSV-RABV-G entry in NTG, AP1G1, AP1B1 and AP1M2 KO cell lines. VSV-RABV-G, VSV particles pseudotyped with rabies virus glycoprotein. n = 3 biological replicates. f. Number of GFP + cells at 6 hpi, from the time-course shown in e. Statistical significance is measured by individual two-sided t-tests comparing GOF lines to NTG, n = 3 biological replicates. For all panels: error bars denote mean ± s.e.m., significance is indicated as: n.s.=not significant, *= P < 0.05, **= P < 0.01, ***=P < 0.001, ****= P < 0.0001.
