Figure 1.

Effect of TSA on cyclin D1 localization and postranslational regulation. A. Asynchronously growing MCF-7 cells were treated with TSA (1 μM) and MG132 (50 μM) alone or in combination and subjected to subcellular fractionation as described in Methods. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, GSK3 and cyclin D1. Immunoblots against mitochondrial Sp1 and Hsp70 (mHsp70) served to ensure efficient subcellular fractionation and mHsp70 also served to monitor equal gel loading. B. Asynchronously growing MCF-7 cells were subjected to subcellular fractionation as described in A. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, Rb, cyclin D1 and mHsp70. C. Asynchronously growing MDA-MB231 were treated as in A. D. Asynchronously growing SKUT-1B cells were treated as in A. E. Asynchronously growing rat KNRK cells were treated as in A. Hsp60 served to ensure efficient subcellular fractionation and also served to monitor equal gel loading.