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. 2003 Jan;23(2):482–492. doi: 10.1128/MCB.23.2.482-492.2003

FIG. 6.

FIG. 6.

EWS/FLI-1 alters C2C12 cell cycle gene expression without interfering with DM-induced cell cycle arrest. Protein lysates were prepared from C2C12 cells expressing EWS/FLI-1 or control cells (neo) cultured in either GM or DM and analyzed by anti-p21 (upper panel), anti-PCNA (middle panel), or anti-cyclin D1 (lower panel) immunoblotting (IB). In the case of the cyclin D1, cells were analyzed in GM and after 3 days in DM. For comparison, protein lysates from the human ES cell lines TC-71, TC-32, and LD were probed with anti-cyclin D1 antibody and shown at the left. (B) C2C12-neo or C2C12-EWS/FLI-1 cells were cultured in GM or cultured for the indicated number of days in DM, labeled with BrdU, and then analyzed in an anti-BrdU-based immunofluorescence assay. The number of cells undergoing DNA replication (BrdU-positive cells) was quantitated by FACS, with the percentage of BrdU-positive cells shown on the right.