Figure 3

NMI does not affect the degradation of IκBα. (A) HeLa cells were transfected with the control or Myc-NMI plasmids. At 24 h posttransfection, the cells were treated with TNF-α (10 ng/ml) for the indicated periods. The cell extracts were subjected to western blotting using the indicated Abs. (B) HeLa cells were transfected with the FLAG-IκBα and Myc-NMI plasmids as indicated. At 24 h post transfection, the cells were treated with TNF-α (10 ng/ml) for the indicated times. The cell extracts were subjected to western blotting using the indicated Abs. (C) HeLa cells were transfected with the control, Myc-NMI or A20 plasmids. At 24 h posttransfection, the cells were treated with TNF-α (10 ng/ml) for the indicated periods. The cell lysates were immunoblotted with the indicated Abs. Statistical analysis results of IκBα expression was in the right panel. Actin was used as the internal control. The results are representative of three independent experiments, and the error bars represent the SD. *p < 0.05, **p < 0.01, ***p < 0.001. (D) NMI binds to IκBα. 293 T cells were transfected with the Myc-NMI plasmid, and the cell lysates were immunoprecipitated with anti-Myc or anti-IκBα Ab and immunoblotted for the indicated proteins.