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Fig. 9. Knockdown of N-cadherin in MCF10A cells decreased cell motility. (A) Wound-healing assays. Phase-contrast micrographs of living cultures of parental MCF10A cells (a-f), MCF10A/ctr KD cells (g-l) and MCF10ANcad cells (m-r) in a wound-healing assays performed in the absence (a-c,g-i,m-o) or presence (d-f,j-l,p-r) of TGF-ß1 (2 ng ml^–1). TGF-ß1 treatment began 2 days before initiation of the assays. Photographs were taken just after the incision (0 hour) (a,d,g,j,m,p) and at 5 hours (b,e,h,k,n,q) and 13 hours (c,f,i,l,o,r) after incision using a 10x objective; scale bar, 50 µm. (B) Motility was quantified by measuring the decrease in the denuded area at 5 hours and 13 hours, and presented as the average decrease in the number of pixels with standard deviation in three independent experiments. TGF-ß1 treatment increased motility in all cell lines tested. In the presence of TGF-ß1, MCF10ANcad cells were significantly slower than parental or control knockdown cells at 5 hours (h) and 13 hours. (C) Transwell motility assays of parental MCF10A cells, MCF10A/ctr KD cells and MCF10ANcad cells in the absence (–) or presence (+) of TGF-ß1 (2 ng ml^–1). TGF-ß1 treatment began 2 days before initiation of the assays. Data are presented as the average number of migrating cells in nine random fields of view in three independent experiments. In the presence of TGF-ß1, MCF10ANcad cells were significantly less motile than parental or control knockdown cells.

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