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. 2021 Nov 25;12:6855. doi: 10.1038/s41467-021-27180-0

Fig. 3. The enhancing effect of T403R in RaTG13 S depends on E37 in ACE2.

Fig. 3

a HEK293T cells expressing indicated ACE2 constructs were infected with VSVΔG-GFP pseudotyped with SCoV-2, RaTG13, or indicated mutant S. Quantification by automatic counting of GFP positive cells. Bars represent the mean of three independent experiments (±SEM). Two-tailed Student’s t-test with Welch’s correction. b Mutations introduced into human ACE2 (upper) and exemplary immunoblot of whole cells lysates (WCLs) of HEK293T cells expressing the indicated ACE2 constructs. Blots were stained with anti-ACE2 and anti-GAPDH and quantified for ACE2 expression (lower panel). Bars represent the mean of three independent experiments (±SEM). c Schematic representation of the ACE2 interaction assay. d Exemplary immunoblots of WCLs of HEK293T cells expressing SCoV-2 S, RaTG13 S, or the indicated mutant. Blots were stained with anti-SCoV-2 S, anti-GAPDH, and quantified for Spike expression. e ACE2 binding using whole cell lysates of HEK293T expressing SARS-CoV-2, RaTG13, or indicated mutant S. Bars represent the mean of three independent experiments (±SEM).