Figure 2

IKKα and IKKβ increase Myc protein level. (A). MCF7 cells were treated with 10 μM Bay11-0782 for 12 hours and then were further treated with 20 ng/ml TNF-α for 30 minutes. The cytoplasmic and nuclear extracts were subjected to Western blot analysis of NF-κB p65 subcellular distribution. Stains of tubulin were used to represent the clearance of cellular fractionation and stains of actin were used as loading control. (B). MCF7 cells were treated with 10 μM Bay11-0782 for 12 hours, and then the whole cell lysates, the cytoplasmic and nuclear extracts were prepared. Equal amount of proteins (15 μg/lane) were subjected to Western blotting analysis. (C). MCF7 cells were transfected with IKKα and IKKβ (both wild-type and kinase-dead mutant). The transfected cells were analyzed by Western blotting of IKKα and IKKβ.(D). The whole cell lysates prepared from indicated cells were subjected to Western blot analysis.