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Figure 4


Fig. 4. Role of MT1-MMP in myotube formation and pericellular fibronectin. (A) Differentiating cells were pulse-treated with the indicated inhibitors during the elongation stage, as indicated in the upper part of the figure. TIMP-1 and TIMP-2 were used at concentrations of 1 µg/ml. The cell fusion ratio was monitored at 144 hours and shown in the relative ratio dealing NT as 100%. The results are presented as the mean ± s.e.m. (n = 6). NT, no treatment; DMSO, solvent for BB94 alone. (B) Endogenous MT1-MMP expression in the C2C12 cells was knocked down by the stable expression of shRNA, using the U6 promoter. A western blot in the upper panel shows a significant reduction of MT1-MMP by shRNA for MT1-MMP (U6MT1), but not for the control LacZ (U6LacZ). In the lower panel, the fusion ratio was monitored at 144 hours and shown in the relative ratio dealing U6LacZ as 100%. The results are presented as the mean ± s.e.m. (n = 6). (C) Cells treated with the indicated inhibitors or with the shRNAs were collected and analyzed by western blotting using antibodies against fibronectin or tubulin.





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