Extended Data Fig. 5: SR-B1 antagonists block SARS-CoV-2pp infection.
From: HDL-scavenger receptor B type 1 facilitates SARS-CoV-2 entry
a, Immunoblot analysis of SR-B1 expression in Huh-7 cells transfected with HA-SR-B1 or siSR-B1 from Fig. 4b–d. b, The bars in the graph represent the RFI from 50 cells per sample from Fig. 4b. n=50 from 3 biologically independent samples and five technical duplication for each sample. P< 0.0001 for siCtrl vs siSR-B1 by two-tailed Student’s t-tests. c, d, The percentage of SARS-2-S-positive cells (c) and the MFI of SARS-2-S (d) on the cell surface were quantified with isotype and SARS-2-S staining from Fig. 4e. n = 3 independent biological experiments. P = 0.0475 for HDL HA-V vs HDL HA-SR-B1 (c), P = 0.0115 for HDL HA-V vs HDL HA-SR-B1 (d) by two-tailed Student’s t-tests. e, Immunoprecipitation analysis in 293 T cells transfected with Flag-SARS-2-S. The approximate molecular weight (kDa) marker positions are indicated to the left of the blot. f, g, Huh-7 cells were preincubated with the indicated concentrations of ITX 5061 (f) or BLT-1 (g) for 48 h, and MTT was added to the cells before OD492 measurements. Signals obtained without compounds were used for normalization. n = 3 independent biological experiments. Nonsignificant (ns) by one-way ANOVA and Bonferroni’s post hoc analysis. h, i, Huh-7 cells preincubated with the indicated concentrations of ITX 5061 (h) or BLT-1 (i) were inoculated with SARS-CoV-2pp in 10% FBS, and pseudotyped viral entry was analyzed by luciferase activity at 48 h after infection. Signals obtained without compounds were used for normalization. n = 3 independent biological experiments. P < 0.0001 for 0 μM ITX 5061 vs 1.56 μM ITX 5061, P < 0.0001 for 0 μM ITX 5061 vs 6.25 μM ITX 5061, P < 0.0001 for 0 μM ITX 5061 vs 25 μM ITX 5061, P < 0.0001 for 0 μM ITX 5061 vs 100 μM ITX 5061 by one-way ANOVA and Bonferroni’s post hoc analysis (h). P = 0.0038 for 0 μM BLT-1 vs 6.25 μM BLT-1, P < 0.0001 for 0 μM BLT-1 vs 12.5 μM BLT-1, P < 0.0001 for 0 μM BLT-1 vs 25 μM BLT-1, P < 0.0001 for 0 μM BLT-1 vs 50 μM BLT-1 by one-way ANOVA and Bonferroni’s post hoc analysis (i). The data are the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.
