Figure 5.

Cytoplasmic sequestration of cyclin D1 in human cancer cell lines. A. Asynchronously growing MCF-7 cells were treated with cycloheximide (Chx) (50 μM) alone and together with MG132 (50 μM) or LMB 10 ng/ml as indicated. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, GSK3, cyclin D1, p21 and Hsp60. B. Asynchronously growing SKUT-1B cell were treated as in A. C. MCF-7 cells were treated for 24 h with the GSK3-specific inhibitor SB216763. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, cyclin D1 and mHsp70.