Research Article

Interaction of Wnt Signaling with BMP/Smad Signaling during the Transition from Cell Proliferation to Myogenic Differentiation in Mouse Myoblast-Derived Cells

Figure 3

Effect of BMP4 and noggin on the myogenic differentiation of C2C12 cells. (a–d) The recombinant proteins were added to the proliferation medium at final concentrations of 5 ng/mL BMP4 and/or 50 ng/mL noggin and cultured in proliferation medium for 3 days, followed by immunostaining with anti-troponin T antibodies (red) and anti-phosphohistone H3 (Ser10) antibodies (green). BMP4 addition inhibited myogenic differentiation in proliferation medium. The number of cells expressing phosphorylated histone H3 was increased by adding noggin but not BMP4. (a) Control; (b) noggin; (c) BMP4; (d) noggin + BMP4. (e–g) The ratios of phosphohistone H3 (e) and troponin T-positive cells (f) to the total cell number were estimated. The number of multinuclear cells expressing troponin T (g) was also estimated to determine the ratio of fused cells among the total cells. Mean + SD ( ), versus control with Student’s -test.
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