Figure S6.

Flow Cytometry for Cell Surface ACE2 Expression and Protein Analysis of RAB7A Protein after CRISPR Targeting, Related to Figure 6

(A) and (B) Flow cytometry gating strategy to quantify cell surface expression of ACE2. (A) Live cells were first gated by the forward and side scatter area, then doublets were excluded by gating with the forward scatter area and width. Viable cells were selected by gating on side scatter area and LIVE/DEAD violet. (B) Gating strategy to determine ACE2+ cells. The gate was position such that < 3% of A549 wild-type and > 85% of A549^ACE2 cells were ACE2 positive. The same gating strategy was applied to all samples. (C) western blot on A549^ACE2 cells perturbed with non-targeting (NT) or RAB7A-targeting guide RNAs and probed with a RAB7A antibody. GAPDH was used as loading control.