JEV infection activates AP-1 via c-Src/PDGFR/PI3K/Akt/MAPKs pathway. (A) Cells were pretreated with U0126 (U0, 1 μM), SB203580 (SB, 1 μM), or SP600125 (SP, 1 μM) for 1 h, followed by stimulation with JEV for 20 min. The isolated RNA samples were analyzed by RT-PCR, using the primers specific for c-Fos, c-Jun, and β-actin. (B) Cells were transfected with an AP-1 promoter luciferase construct together with a β-galactosidase plasmid, pretreated with U0126 (U0, 1 μM), SB203580 (SB, 1 μM), or SP600125 (SP, 1 μM) for 1 h, and then incubated with JEV for 30 min. AP-1 promoter activity was normalized to that of β-galactosidase activity. (C-E) Cells were pretreated with AG1296 (AG, 10 μM), PP1 (10 μM), or LY294002 (LY, 30 μM) for 1 h, and then infected with JEV (moi = 1) for (C) 5, (D) 10, or (E) 30 min. The cell lysates were analyzed by western blot using an anti-phospho-p42/p44 MAPK, anti-phospho-p38 MAPK, anti-phospho-JNK1/2, or anti-GAPDH antibody. Data are expressed as mean ± S.E.M. for five independent experiments. #P < 0.01, as compared with the cells exposed to JEV alone.
Yang et al. Journal of Neuroinflammation 2012 9:12 doi:10.1186/1742-2094-9-12