Figure 3

JEV infection induces MMP-9 expression through c-Src/PDGFR in RBA-1 cells. (A) Cells were pretreated with AG1296 (10 μM) or PP1 (10 μM) for 1 h, and then infected with JEV (moi = 1) for the indicated time intervals. The cell lysates were analyzed by western blot using an anti-phospho-PDGFR or anti-GAPDH antibody. (B, C) Cells were pretreated without or with AG1296 (AG, 10 μM) or PP1 (10 μM) for 1 h, and then incubated with JEV (moi = 1) for the indicated time intervals (B) or 5 min (C). Cell lysates were subjected to immunoprecipitation using an anti-c-Src antibody. The immunoprecipitates were analyzed by western blot using an anti-c-Src, anti-phospho-c-Src, or anti-phospho-PDGFR antibody. (D, E) Cells were pretreated with AG1296 or PP1 for 1 h or transfected with PDGFR siRNA, followed by incubation with JEV for 16 h. Cell lysates were analyzed by western blot using an anti-PDGFR or anti-GAPDH antibody. The conditioned media were used to determine MMP-9 expression by gelatin zymography. Data are expressed as mean ± S.E.M. for five independent experiments. ^#P < 0.01, as compared with cells exposed to JEV alone [D (left panel), E]. ^#P < 0.01, as compared with cells transfected with scrambled siRNA exposed to JEV (D, right panel).