Fig. 7.

GBP5 stimulates the expression of COX-2 during IAV infection. a A549 cells were transfected with 2 μg of GBP5 expression plasmid for 24 h, then the cells were infected with IAV (MOI = 1) and harvested at 24 hpi. Relative mRNA levels of the indicated genes were measured by qRT-PCR. b A549 cells were transfected with vector or GBP5 for 24 h, then the cells were infected with IAV (MOI = 1) and harvested at the indicated time. Relative levels of COX-2 mRNA were measured by qRT-PCR. c A549 cells were transfected with the GBP5 expression plasmid in a dose gradient for 24 h, then the cells were infected with IAV (MOI = 1) and harvested at 24 hpi. The relative level of COX-2 mRNA was measured by qRT-PCR and the protein level was measured by Western blot. d 293T cells were transfected with luciferase reporter plasmids containing the wild-type COX-2 promoter or mutant COX-2 promoter, along with IAV (MOI = 1) infection. Luciferase activities were measured in each sample after serum starvation for 24 h, and were compared to those obtained from control cells transfected with empty vector. e A549 cells were transfected with the GBP5 expression plasmid and then infected with IAV (MOI = 1) for 12 h and ChIP assays were performed using 2 μg of anti-CREB1. Normal rabbit IgG was used as a control. Immunoprecipitated DNA or control DNA was collected and amplified using specific primers for the CREB1 binding site in the COX-2 promoter. Bar graphs represent means ± SD; n = 3 (** p < 0.01; * p < 0.05).