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

FIGURE 1.

FIGURE 1.

Inhibition of Matrigel invasion induced by N6L and HB-19 treatment is associated with TIMP-3 augmentation in culture media. A and B, MDA-MB-435 cells seeded in the upper compartment of modified Boyden chambers containing 20 μg of Matrigel in FBS-free medium and incubated for 24 h for invasion in the presence or not of 10 μm N6L or HB-19. DMEM supplemented with 5% FBS was placed in the lower chamber as chemoattractant. Invading cells were visualized by May-Grünwald/Giemsa staining and counted using ImageJ software (supplemental Fig. 3). A, cell number ± S.E. (error bars; n = 4). *, p < 0,05; ***, p < 0,001 statistically significant compared with control. B, representative images of membrane after Matrigel invasion by cells. Scale bars, 50 μm. C–E, MDA-MB-435 cells were treated or not for 48 h at 37 °C with 10 μm N6L or HB-19 in FBS-free medium. Sample loading was normalized using protein concentration of cells lysates. C, zymogram for MMP detection in the media conditioned by MDA-MB-435 cells. D, reverse zymogram for TIMP detection in the media conditioned by MDA-MB-435 cells. E, Western blot analysis for TIMP-3 expression in the media conditioned by MDA-MB-435 cells.