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. 2009 Apr 7;17(6):1064–1072. doi: 10.1038/mt.2009.67

Figure 2.

Figure 2

Muscle differentiation markers expressed in MyoD-hMADS cells. (a) Comparative expression by RT-PCR between WT- and MyoD-hMADS cells after 3 and 7 days in myogenic differentiation medium. Many markers could be amplified at day 7 for MyoD-hMADS cells. PCR was run for 30 cycles, which gave sufficient amplification signal for MyoD-hMADS cells at day 7 without any amplification for other cDNAs under comparison. All the primers were human sequence specific. (b) Immunofluorescence analysis of MyoD-hMADS cells in myogenic differentiation medium. The muscle proteins desmin, nestin, skeletal myosin, and dystrophin were detected using specific antibodies revealed with Rhodamin fluorochrome (red). Nuclei were counterstained with Hoechst 34580 (blue) to detect multinucleated myotubes. Note the presence of mononucleated hMADS cells also expressing desmin beside the desmin positive myotubes. Skeletal myosin is detected with an anti-myosin heavy chain. Dystrophin could be observed in some myotubes at its expected membrane localization. Bar = 20 µm. cDNA, complementary DNA; RT-PCR, reverse transcriptase-PCR; WT-hMADS, human multipotent adipose-derived stem.