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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Mol Cancer Res. 2013 Nov 7;12(1):69–81. doi: 10.1158/1541-7786.MCR-13-0300

Figure 4. NEDD9 depletion decreases MMPs activity, but does not affect MMPs expression or secretion.

Figure 4

(A). WB of total MMPs expressed (WCL) and secreted (SFM) by indicated cells, 48h post shRNA expression. Schematic outline the MMP microarray. (B). Quantification of MMPs and TIMPs expression, n=3, one-way ANOVA for all pairs p is non-significant; p<0.05 for MMP13 (shCon/shN4). (C). Quantification of gellatinase/collagenese activity in SFM conditioned by cells using EnzChek Gellatinase/Collagenese assay. Clostridium Collagenase, 0.1U/ml, is positive control, p<0.05 (shCon/shN1 or /N4), linear regression analysis. (D). Representative image of -gel zymography, SFM conditioned by MDA-MB-231-shCon, -shNEDD9 cells, WB of MMP2 and MMP9. (E). Quantification MMP2 and MMP9 activity as in (D) normalized to total level of MMP2 and MMP9, +/− S.E.M; MMP9: p=0.007 (*) shCon/N1, or /N4 and ns - shN1/N4, one-way ANOVA. MMP2: p=0.00031, shCon/N1, /N4, ns -shN1/N4, one-way ANOVA. (F). MMP14-specific fluorogenic substrate assay, at 328/400mn. Data plotted as curves described by equation for one site binding kinetics, fitted to each data set with the R2 of 0.8, Extra sum-of-squares F test, p<0.0001.