Fig. 7: TGFβ signalling regulates a mechanically induced tumour promoting phenotype in normal myoepithelial cells.
a Representative images of integrin β6 (green) and fibronectin (FN, magenta) by immunofluorescent staining in stretched normal myoepithelial cell line; N-1089 treated with a control (CTLAB) or TGFβRII (RIIAB) blocking antibody. Nuclei were counterstained with DAPI (blue). Scale bar, 20 μm. b Immunoblots of integrin β6 and fibronectin in stretched normal myoepithelial cell lines generated as in (a). HSC70 serves as loading control. Images are representative of three experiments. c, d Bar graphs showing qRT-PCR analysis for ITGB6 (c) and FN1 (d) using RNA isolated from stretched normal myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 3 biological replicates, repeated 3 times; ****P < 0.0001 (two-tailed t-test). e Invasion assay for MDA-MB-231 (231) and MCF-7 cells using conditioned media (CM) isolated from stretched normal myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 3 biological replicates, repeated 4 times; MDA-MB-231 ***P = 0.0001 and MCF-7 ****P < 0.0001 (two-tailed t-test). f, g Bar graphs showing qRT-PCR analysis for MMP9 (f) and MMP13 (g) using RNA isolated from stretched normal myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 3 biological replicates, repeated 3 times; ***P = 0.0001 (f) and **P = 0.0041 (two-tailed t-test).
