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. 2012 Oct 29;287(52):43685–43693. doi: 10.1074/jbc.M112.380402

FIGURE 6.

FIGURE 6.

Suppression of N6L inhibitory activity on cell invasion using silencing of TIMP-3 in MDA-MB-435 cells. MDA-MB-435 cells were transfected for 48 h I 100 nm TIMP-3 siRNA or GFP siRNA as negative control or without siRNA as transfection control (L2000) using Lipofectamine 2000 protocol. A, mRNA TIMP-3 expression performed by RT-PCR. B, TIMP-3 protein expression analysis performed by Western blotting. C, TIMP-3 release from silenced cells or not for TIMP-3 in culture media after 10 μm N6L treatments and analyzed by Western blotting. D and E, after transfection, cells invasion evaluated in modified Boyden chambers as described in Fig. 1. D, cell number ± S.E. (error bars; n = 4). **, p < 0.01; ***, p < 0.001 statistically significant compared with control. E, representative pictures of membrane after Matrigel invasion by cells. Scale bars, 200 μm.