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. 2008 Jul 25;283(30):21230–21241. doi: 10.1074/jbc.M710525200

FIGURE 4.

FIGURE 4.

Effect of forced ca-MRTF-A expression on myogenic differentiation. A, experimental procedures are presented schematically at left. Stable C2C12 transfectants (C2C12-tet-off-FALG-ca-MRTF-A) were cultured under the conditions described in the legend for Fig. 1D. Cells (5-day cultures in DM) were stained with an anti-MHC antibody (right). Magnification is ×100. B and C, expression levels of myogenic differentiation markers and myogenic inhibitors are shown. B, whole-cell extracts from C2C12 cells cultured under the indicated conditions (5 days in DM) were subjected to immunoblotting using the indicated antibodies. C, total RNA from C2C12 cells cultured under the indicated conditions (5 days in DM) was analyzed by RT-PCR as described in the legend for Fig. 3. Lanes 1-3 (B and C) correspond to those in A. D, C2C12 cells were transfected with siRNAs against Id3 (Id3 siRNA1 and -2) and were cultured in GM for 2 days. The whole-cell extracts were subjected to immunoblotting using the indicated antibodies. The cell morphology and expression of MHC were also analyzed by staining with an anti-MHC antibody (red) and Hoechst 33258 (blue). Magnification is ×200.