Figure 2From: The cyclin D1 proto-oncogene is sequestered in the cytoplasm of mammalian cancer cell linesCyclin D1 localization in human cancer cells and murine tissues. A. Asynchronously growing U2OS, HeLa and KNRK cells were subjected to subcellular fractionation as described in Methods. Cell lysates and insoluble fractions were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, GSK3 and cyclin D1. B. Indicated mouse tissues were subjected to subcellular fractionation as described in the Methods section. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against cyclin D1. Immunoblots against Rb and Hsp60 served to ensure efficient subcellular fractionation. C. Asynchronously growing MCF-7 cells were subjected to proteomic fractionation. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, GSK3 and cyclin D1. Immunoblots against Sp1 and Hsp60 served to ensure efficient subcellular fractionation. D. Asynchronously growing MCF-7 cells were subjected to subcellular fractionation using an in-house protocol. Cell lysates were separated by 4–20 % SDS-PAGE and immunoblot analysis was done using antibodies against Sp1, GSK3 and cyclin D1.Back to article page