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. 2010 Jun 3;6(6):e1000930. doi: 10.1371/journal.ppat.1000930

Figure 6. NOX2 is required for RIG-I-mediated regulation of IFNβ and IFIT1 genes.

Figure 6

(A and B) A549 were cotransfected with the ISG56-pGL3 firefly luciferase and the pRL-null renilla luciferase (internal control) reporter constructs. Cells were then pretreated with 3mM Tempol, 10µM DPI, 1mM Apo (white bars) or the corresponding vehicle (black bars) before being left unstimulated (NS) or transfected with poly I:C. (C) A549 were transfected with CTRL (black bars) or NOX2 (white bars) RNAi and were further cotransfected with the IFNβ- or ISG56-pGL3 and pRL-null reporter constructs and transfected with poly I:C for 8h. (A–C) Luciferase activities were measured and expressed as described in Figure 1. (*, p<0.05;**, p<0.01; ***, p<0.001; mean ± SEM of triplicates). (D) A549 were transfected with CTRL (black bars) or NOX2 specific (white bars) RNAi and left unstimulated or transfected with poly I:C for 3h. mRNA levels of IFNβ and ISG56 were analyzed by real-time PCR as described in Figure 1 after normalization to S9 mRNA used as a reference gene. NOX2 immunoblot was performed as described in Figure 2A. (*, p<0.05; ***, p<0.001; mean ± SEM of independent triplicates).