Figure 4: Validation of key genes identified in CRISPR-Cas9 whole genome screening using the one vector system. A) ACE2 staining on the ACE2 KO cells using AF488 conjugated anti-ACE2 antibody (dark blue) which is similar to unstained (magenta) versus WT ACE2 positive cells (light blue). B) ACE2 staining on the ACE2-293T-CRE reporter cells following transduction with the ACE2 targeting sgRNA (dark blue) relative to WT (light blue) and unstained (magenta). C-F) Effect of mutants on the entry of mNG and Cre reporter viruses. The error bar represents three biological replicates. One-way ANOVA and Šídák’s multiple comparisons test were performed to do statistical analysis between the WT and the mutants. Bar shows mean ± SD (n= 3). * Denotes p<0.05 for comparison between WT and the mutants.