Fig. 4.

Expression and biological significance of the cyclin D1b isoform in EwSa. (A) RT-qPCR analysis of cyclin D1 transcripts in total RNA from the indicated samples. Each dot represents a sample. The average ± SEM is also indicated. CLs, cell lines. (B) Total proteins from various EwSa cell lines, tumors, and A673 cells transfected with the indicated siRNAs were analyzed by Western blot using antibodies against D1a, D1b, and actin. (C and D) Cytosolic (C) and nuclear (N) proteins from EW7 and A673 cells were analyzed by Western blot using the indicated antibodies. A lower exposure of the EW7 cytosol hybridized with the DCS6 antibody is shown. (E) A673 cells transfected for 3 days with the indicated siRNAs were analyzed for cell growth.