Figure 1.

JEV-induced fluctuation of HMGB1 in HBMECs. HBMECs were infected with JEV-P3 at an MOI of 1, and total cell protein and RNA samples were collected at the indicated times to measure JEV replication in HBMECs by real-time PCR (A) and Western blotting using an anti-JEV-E protein monoclonal antibody (B). HMGB1 expression was measured by real-time PCR (C) and Western blot (D) at the indicated times during JEV infection. JEV free cells were served as control. (E) Total cytoplasmic and nuclear proteins were extracted from JEV-infected HBMECs at 0 h (Control), 6 h, 12 h, 24 h, and 48 h postinfection. HMGB1 protein expression was measured by Western blotting, with beta-actin as the internal control for protein integrity and Lamin A/C was assessed in the nuclear extract, and quantitatively analyzed as the fold change relative to the control. (F) HBMEC culture supernatant was collected at indicated times after virus infection (24 h, and 48 h). Supernatant HMGB1 was measured by Western blotting and quantitatively analyzed as the fold change relative to the control (JEV-free cell culture supernatant). The experiments are repeated at least three times. The data are expressed as the means ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001.