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Figure 7


Fig. 7. Downregulation of M-CAM expression in C2C12 cell line by RNAi. (A,B) Negative control (no M-CAM knockdown). (C,D) M-CAM oligonucleotide 20. (E,F) M-CAM oligonucleotide 38. (A,C,E) Immunofluorescence images of each culture stained for M-CAM expression. Note the reduction of M-CAM expression in C and E compared with A. (B,D,F) Cultures at day 7 after addition of differentiation medium stained for expression of myosin heavy chain (green) and with DAPI (blue) to visualize the nuclei. Magnification, 400x (A,C,E); 100x (B,D,F). (G) Quantitative real-time PCR of M-CAM RNA in control, oligo 20- and oligo 38-treated C2C12 cultures. Cultures treated with oligo 20 yielded a 60% reduction in M-CAM mRNA expression, whereas cultures treated with oligo 38 yielded an 87% reduction of M-CAM mRNA. (H) Fusion index determined for each culture after differentiation was induced for 3, 7 and 12 days. The fusion index was calculated as the number of nuclei within myotubes divided by the total number of nuclei.





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