Fig. 5.

HP-BCD treatment inhibits virus RNA and protein expression and reduces SARS-CoV-2-mediated CPE. Vero cells were treated with 20 mM HP-BCD for 1 h. The cells were washed, and mock-treated (control) or infected with SARS-CoV-2 (gamma isolate; MOI = 0.1) for 48 h. A-C) Expression and distribution of dsRNA and cholesterol were evaluated by staining the cells with J2 antibody and filipin, followed by fluorescence microscopy analysis. Representative images are demonstrated in (A; scale bar = 20 μm), with a zoom of infected cell depicted in (B); mean fluorescence intensity (MFI) and frequency of J2 positive staining (% dsRNA+) of 5 randomly selected fields, with a total of about 500 cells, were calculated using ImageJ software and are represented in (C) and (D), respectively. E-G) Calu-3 cells were lysed and the expression of Spike protein was analyzed by western blotting. A representative blot is demonstrated in (E) and the expression level of whole S (F) or S1 (G) was calculated based on β-actin loading control staining, using ImageJ software. Statistical analyses were performed by unpaired t-test; * represents p < 0.05; ***p < 0.001; insert numbers indicate the percentage inhibition in relation to untreated cells.