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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 6. Inhibition or depletion of MMP14 or TIMP2 recapitulates invasion deficiency of shNEDD9 cells.

Figure 6

(A–B). WB of WCL of MDA-MB-231-shCon, -shNEDD9 cells treated with siCon, siMMP14 or siTIMP2, using anti-NEDD9, -TIMP2 and -a-tubulin antibodies. (C). Boyden Chamber invasion assay, cells as in (A–B); plotted as relative fluorescence units (RFU), one-way ANOVA *p<0.0001 (shCon/siCon, shN1/siCon, or shN4/siCon), ns -shN1/N4-siCon and shCon/siMMP14 or shCon/siTIMP2; p<0.005 shCon/siCon, shCon/siMMP14 or shCon/siTIMP2; (D). Boyden Chamber invasion assay of MDA-MB-231-shCon cells with increased amount of TIMP2. MDA-MB-231-shN4 cells used as a reference control. Results plotted as RFU, n=3. One-way ANOVA *p<0.0001 shCon-0nM/shCon-10nM or more, ns: shN4/shCon-10nM. (E). WB of NEDD9 rescue in MDA-MB-231-shCon, -shNEDD9 cells with empty vector or cDNA-NEDD9 construct, using anti-NEDD9, -GAPDH antibodies. (F). FACS of MMP14 surface staining as in (E); one-way ANOVA *p<0.0001 empty vector-shCon/shN1 or /N4; ns cDNANEDD9-shCon/shN1 or /N4). (G). Boyden-Chamber assay, cells as in (E–F). Data plotted as RFU, one-way ANOVA *p<0.0001 (shCon-empty/shCon-NEDD9, shN1-empty/shN1-NEDD9, shN4-empty/shN4-NEDD9); p<0.005 (shCon-empty/shN1-empty or -shN4-empty); ns (shCon-NEDD9/shN1-NEDD9 or /shN4-NEDD9).