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 7

Effects of BMP4 and noggin on β-catenin localization with or without Wnt3a and Wnt4. (a–i) Recombinant adenoviruses were used to express Wnt3a, Wnt4, or eGFP (control) uniformly in C2C12 cells. Cells were infected 48 hours earlier with an MOI of 400, and the recombinant proteins were added as shown in Figure 3 with final concentrations of 5 ng/mL BMP4 and/or 250 ng/mL noggin. Two hours after BMP4 addition, immunostaining for β-catenin was carried out for counting. (j) Comparison of the β-catenin signals after adenovirus-mediated expression of Wnt3a and eGFP, after pixel intensity analysis for immunohistochemical signals with BZ-II Analyzer, shown in abscissa for signal intensity and ordinate for frequency. Wnt3a intensified signals to a higher level. (k and l) Comparison of the β-catenin signals after adenovirus-mediated expression of Wnt3a, Wnt4, and eGFP, in the presence or absence of BMP4 and/or noggin. Immunostaining for β-catenin was analyzed by digitizing the signals with BZ-II Analyzer for comparison. Typical results are shown. (k) Without BMP4; (l) with BMP4.
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