Figure 2. Flow cytometric analysis of pseudotyped lentiviral vectors on stable HEK293T/17-hACE2+TMPRSS2+.
(A) Gating strategy starting with live (eF780) vs. FSC-A to identify live cells, followed by SSC-A vs. FSC-A to identify HEK293T/17 cells, and by SSC-W vs. FSC-A for singlets identification. (B) Transiently produced VSV-G LV, SΔERR CoV-2 LV, and S(D614G)ΔERR CoV-2 LV were serially diluted from 100 µl in a 4-fold dilution for six points and added on control HEKT293T/17 cells (blue population) and CoV-2 S permissive HEK293T/17-hACE2+TMPRSS2+ cells (red population) in the presence of 8 µg/ml of polybrene. Staggered histograms of transduced populations are presented. (C) Graph representing viral titers of the three pseudotyping glycoproteins quantified by an infectivity assay on HEK293T/17 and HEK293T/17-hACE2+TMPRSS2 cells. Data presented are ±SD of duplicate determinations.