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. 2021 Aug 5;142:112011. doi: 10.1016/j.biopha.2021.112011

Fig. 3.

Fig. 3

Effect of GB-2 on interaction between ACE2 and SARS-CoV-2 Spike (K417N, E484K, N501Y or L452R). (A, C, E, G) Flow cytometry analysis of ACE2-Spike protein binding. 293T cells with pCEP4-myc-ACE2 plasmid were incubated with RBD (N501Y(A), K417N(C), E484K(E) or L452R(G))-sfGFP-containing medium and co-stained with fluorescent anti-MYC Alexa 647 to detect surface ACE2 by flow cytometry. During analysis, the top population were chosen from the ACE2-positive population. Then, two subsets of the ACE2-positive population were collected: the top population (nCoV-S-High sort, red gate) and the bottom population (nCoV-S-Low sort, green gate) based on the fluorescence of bound RBD (N501Y(A), K417N(C), E484K(E) or L452R(G))-sfGFP relative to ACE2 surface expression. (B, D, F, G) Quantitative results of nCoV-S-High sort and nCoV-S-low sort, which were presented as ratio compared with blank, in the top population or ACE2-positive population. All the results are representative of at least three independent experiments. (Error bars=mean±S.E.M. Asterisks (*) mark samples significantly different from control group with p < 0.05).