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Fig. 3 | Molecular Brain

Fig. 3

From: Development of a novel peptide to prevent entry of SARS-CoV-2 into lung and olfactory bulb cells of hACE2 expressing mice

Fig. 3

TAT-peptides inhibit SARS-CoV-2 S protein entry into HEK293T cells. A Representative fluorescent microscopy images are shown. HEK293T cells transiently transfected with hACE2 and TMPRSS2 were pre-incubated with vehicle, ACE2 antibody, TAT alone or related TAT-peptides (10 µM), and inoculated with pseudotyped VSV expressing SARS-CoV-2 S protein. Pseudotyped VSV contains a GFP reporter gene. ACE2 antibody was used as a positive control. Scale bar: 100 µm. B TAT-ACE2-1, TAT-ACE2-2, TAT-ACE2-3, TAT-S1/S2, or TAT-S2 inhibit the SARS-CoV-2 S protein entry into HEK293T cells. The fluorescent intensity of GFP in live cells was quantified using a Synergy H4 plate luminometer. Data are shown as mean ± SEM, and presented as the percentage of the control (vehicle) samples. n = 8 – 14 samples for each group, one-way ANOVA test followed by Dunnett’s post hoc test. *p < 0.05, ***p < 0.001, ****p < 0.0001 as compared to control (vehicle) samples. #p < 0.05, ###p < 0.001 as compared to TAT-ACE2-2 samples. C Analysis of EC50 of TAT-ACE2-2 peptide for inhibiting VSV-ΔG-GFP-SARS-CoV-2 pseudovirus entry into HEK 293 T cells transfected with hACE2 and TMPRSS2. Eight dilutions ranging from 3 nM to 3 µM were assessed. D TAT-ACE2-2 inhibits the binding of purified S protein and ACE2. The RBD domain of SARS-CoV-2 S protein was coated onto a 96-well plate, and the same concentration of purified ACE2 together with various concentrations of TAT-ACE2-2 were simultaneously added into each well. ACE2 binding to S protein was detected using anti-ACE2 antibody and HRP-conjugated secondary antibody; TMB one-step substrate reagent was added to visualize HRP. Data are shown as mean ± SEM, and presented as the percentage of the control samples (no TAT-ACE2-2 added, 0 mM). n = 3 samples for each group; one-way ANOVA test followed by Dunnett’s post hoc test. *p < 0.05, ****p < 0.0001 as compared to control (0 mM) samples

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