Figure 6.

A role for IKKβ and IRF5 in the increased transcription of NKG2DL after VVΔE3L infection. A, fibroblasts expressing the NPro gene of BVD virus were either left uninfected or infected with the VVΔE3L virus. qRT-PCR was used to assay MICA and ULBP2 expression (4 experiments). B, fibroblasts were either left uninfected or infected with the indicated virus in the presence or absence of 5 μm BI605906 that inhibits IKKβ. qRT-PCR was used to assay MICA, ULBP2, and VVA17 expression (3 experiments). C, Western blot analysis of IRF5 expression in cytosolic and nuclear lysates of fibroblasts infected with the indicated viruses, Lamin A was used as a fractionation control (2 experiments). D, 293T cells were co-transfected with Firefly luciferase reporter plasmid containing either wt or mutant ULBP2 promoter, Renilla luciferase, and an expression vector for the indicated IRF5 isoform. After 24 h, cells were lysed and the Renilla and Firefly luciferase activities were measured by the dual luciferase assay (9 experiments). The data were normalized using the Renilla luciferase levels. Data are presented ± S.D. Statistical significance was determined using one-way ANOVA (*, p < 0.05; **, p < 0.01; ***, p < 0.001; no bars, S.D. values smaller than symbol used; no symbol means not statistically significant, p > 0.05).