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. 2015 Jan 14;6(1):77–88. doi: 10.1080/19491034.2015.1004947

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Figure 7. — Stable knockdown of nesprin-2 in C2C12 cells reduces efficiency of myoblast fusion without affecting MyoD expression. A. Immunofluorescence of nesprin-2G (green) and staining of actin (red) and nuclei (blue) in C2C12 expressing control shLuciferase (shLuc) and nesprin-2 shRNA (shN2) 4 d after induction of differentiation. Note reduction of nesprin-2 levels and formation of multinucleated myotubes. B, C. Fusion index (ratio of number of nuclei in myotubes to total number of nuclei multiplied by 100) and number of nuclei per myotube in C2C12 cells differentiated for 4 d. At least 2000 total nuclei were count for each condition. D) MyoD immunofluorescence (green) and actin (red) staining of C2C12 cells differentiated for 4 d. E. Fusion index of C2C12 myoblasts expressing control Luciferase (shLuc) and nesprin-2 (shN2) shRNAs induced to differentiate for different times. Statistical analysis was performed to compare fusion index of shN2 to shLuc controls. Bars in A and D, 10 μm. Error bars in B,C,E are SD from 3 experiments. ns, not significant; **, P < 0 .01 by t-test.