FIG. 2.

The MVA/5.2kb virus inhibits IκBα degradation and NF-κB activation in 293T and RK13 cells. (A and B) 293T cells (A) and RK13 cells (B) were either mock infected (UN) or infected at an MOI of 10 with either the WR, MVA, or MVA/5.2kb virus. At 8 h postinfection, cells were collected and lysed in CE buffer. Equal amounts of cytoplasmic extracts were analyzed on sodium dodecyl sulfate-12% polyacrylamide gel electrophoresis and then by Western immunoblotting with anti-IκBα antisera. (C and D) 293T cells were transfected with pRL-null and pNF-κBluc (C) and RK13 cells were transfected with pRL-TK and pNF-κBluc (D). For both cell types, at 24 h posttransfection cells were either mock infected (UN) or infected at an MOI of 10. Eight hours postinfection, cells were collected and lysed, and firefly and sea pansy luciferase activities were measured. The luciferase assays were performed in triplicate, and each bar represents the averaged data from one representative experiment. The relative fold changes were determined by normalizing the ratios of firefly luciferase activity in virus-infected cells to sea pansy luciferase activity in each virus-infected cell group to the value obtained for uninfected cells (see Table 1 for raw data).