Figure 6

IKKs-Myc pathway is inducible. (A) MCF7 cells were treated with 10 μM Bay11-0782 alone, or 200 nM doxorubicin in the presence or absence of 10 μM Bay11-0782 for 24 hours (Dox, Bay, and Bay/Dox), or 3-hr short-term pulse followed by 24 hours release in the presence or absence of Bay11-0782 (Dox3-R, Bay/Dox3-R, and Bay/Dox3-R/Bay). Whole cells lysates were subjected to Western blot analysis. (B) MCF7 cells were treated 200 nM doxorubicin with or without 10 μM Bay11-0782 for 24 hours. The cytoplasmic and nuclear lysates were subjected to Western blotting. (C) MCF7 cells were treated with 200 nM doxorubicin in the presence or absence of 10 μM Bay11-0782 for 3 hours. The cells were then seeded into soft-agar in complete medium with or without 10 μM Bay11-0782 for two weeks. Each experiment have three 60 mm dishes and each data represents mean ± SD calculated from two independent experiments. (D) MCF7 cells were treated as shown, and then the ability of cells to penetrate matrigel was determined. Each experiment have three trans-wells and each data represents mean ± SD calculated from two independent experiments.