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Fig. 8. TGF-ß1-mediated EMT in MCF10A cells. (A) MCF10A cells in the absence (a) or presence (b) of TGF-ß1 [2 ng ml^–1 for 1 day (d)] were stained for F-actin. Photographs were taken using a 63x oil objective; scale bar, 10 µm. (B) Phase-contrast micrographs of TGF-ß1-treated MCF10A cells (t=0-3 days). Photographs were taken of living cells using a 10x objective; scale bar, 50 µm. (C) TNE extracts of TGF-ß1-treated parental MCF10A cells (t=0-3 days; lanes 1-4) control knockdown cells (ctr KD cells) (t=0-3 days; lanes 5-8), N-cadherin knockdown cells (Ncad cells) (t=0-3 days; lanes 9-12) and untreated N-cadherin overexpressing cells (Ncad cells) (lane 13) were immunoblotted for N-cadherin (regular and longer exposure), E-cadherin, GAPDH and fibronectin. The slower migrating bands in the N-cadherin immunoblot are pro-region-containing precursor forms of N-cadherin. (D) Quantification of N- and E-cadherin levels using normalization with GAPDH. (E) MCF10ANcad in the absence (a,c) or presence (b,d) of TGF-ß1 (2 ng ml^–1 for 2 days) were co-stained for paxillin (a,b) and F-actin (c,d). Photographs were taken using a 63x oil objective; scale bar, 10 µm.

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