Fig. 3.

HP-BCD does not affect the expression of SARS-CoV-2 receptors. ACE-2-expressing HEK-293 cells or Calu-3 cells were treated with 20 mM HP-BCD for 1 h. The expression of ACE2 in intact nonpermeabilized cells was analyzed by flow cytometry, whereas total expression of ACE2 and CD147 were evaluated in the cell lysates by western blotting. Representative histograms of ACE2 staining and flow cytometry analysis are depicted in (A) and (F); and the MFI average obtained from independent experiments represented in (B) and (G). For the western blotting analysis, the membranes were stained with anti-ACE2, anti-CD147, and anti-β-actin, as a loading control. Representative blots are demonstrated in (C) and (H); the expression level of ACE-2 (D, I) and CD147 (E, J) were estimated using ImageJ software and normalized according to β-actin expression. Three experiments were run with Calu-3 cells and statistical analyses were performed by paired t-test.