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Fig. 4. Sca-1 downregulation blocks myogenesis. Untransfected, control C2C12 cells (C) and cells expressing Sca-1 antisense (AS) were grown under proliferation conditions and in differentiation medium for 5 days (as described for Fig. 3). They were then metabolically labeled with BrdU and stained with DAPI to identify nuclei, with antibody against BrdU (red) to detect DNA synthesis, and with antibodies against Myf5 (green), MyoD (green) and myogenin (green) as markers of myoblast differentiation. Bar, 50 µm. (A) Whereas proliferating (Pro) control and antisense cells expressed Myf5 and MyoD, and myogenin was expressed under differentiation conditions in both cell lines, expression of the early myogenic marker Myf5 persisted in Sca-1 antisense cells under differentiation conditions. Quantitative analysis provided in text. (B) Wild-type cells were negative for Myf5 and BrdU after 5 days in differentiation medium, whereas Sca-1 antisense cells grown in differentiation medium for 5 days were positive for both Myf5 and BrdU. Quantitative analysis provided in text.
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