FIGURE 5.
Cytosolic calcium outward induced by Evans blue is required for HBV capsid assembly. (A) HuhDhNTCP was treated with compounds after infection for 6 days, supernatant HBV and total HBV DNAs were detected by real-time PCR. (B–D) Drug toxicity and antiviral effects of long-term (6 days) treatment of Evans blue on HepAD38 cell line. The antiviral effects of Evans blue treatment were determined by HBeAg, HBsAg, total DNA, and supernatant HBVs. (E,F) Cytosolic calcium levels in HepAD38 cells treated with Evans blue alone (E) or with Evans blue and 10 μM thapsigargin (F) were determined using a calcium dye. (G) HepAD38 cells plated in 6-well plates were treated with different concentration of Evans blue for 6 days, and the HBV core protein levels were detected by Western blot, β-actin was used as an internal reference. A particle gel assay followed by western blot was conducted to measure the capsid levels.