Figure 1.

Regulation of COX‐2 expression by EV71 infection occurs through CREB. (A) Cells were incubated with H89 for 1 h, followed by infection with EV71 for 12 h. (B) Cells were transfected with CREB siRNA, and were incubated with EV71 for 12 h. (C) Cells were pretreated with H89 for 1 h before incubation with EV71 for 16 h. (D) Cells were pretreated with H89 for 1 h, followed by incubation with EV71 for 1 h. (E) Cells were transfected with COX‐2‐luc reporter gene, pretreated with H89 for 1 h, and then incubated with EV71 for 16 h. (F) Cells were transfected with the wildtype COX‐2 promoter and CRE mutation COX‐2 promoter, prior to incubation with EV71 for 16 h. (G) Cells were incubated with EV71 for the indicated time. (H) Cells were pretreated with H89 for 1 h before stimulation with EV71 for 30 min. The cell lysates were subjected to Western blot analysis using an anti‐CREB (B, G, H), anti‐COX‐2 (A, B), and anti‐p‐CREB (G, H) antibody. The medium was analyzed for PGE₂ release (C). The COX‐2 mRNA expression was analyzed by RT‐PCR (D). The COX‐2 promoter activity was determined in the cell lysates (E, F). Data are expressed as mean ± SEM of at least three independent experiments. *P < 0.05; ^# P < 0.01 as compared with the cells exposed to EV71 alone (A–E, H). ^# P < 0.01 as compared with the cells transfected with the wild‐type COX‐2 promoter in response to EV71 (F). *P < 0.05; ^# P < 0.01 as compared with the basal level (G).