Fig. 3: Integrin β6-positive myoepithelial cells promote breast cancer cell invasion by fibronectin-mediated activation of TGFβ signalling.
a Migration assay for myoepithelial cell line; β6-1089 transfected with a control (CTL) or fibronectin-targeted (FNKD) siRNA, to BSA and LAP (error bars, +s.e.m). n = 3 biological replicates, 3 technical replicates; ****P < 0.0001 (two-tailed t-test). b Adhesion assay for myoepithelial cell lines generated as in (a) to LAP (error bars, +s.e.m). n = 3 biological replicates, 3 technical replicates; ***P = 0.0001 (two-tailed t-test). c Bar graph showing qRT-PCR analysis for TGFB1 using RNA isolated from myoepithelial cell lines generated as in (a). n = 3 biological replicates, 3 technical replicates; ****P < 0.0001 (two-tailed t-test). d Immunoblots of phospho-SMAD2 (pSMAD2) and SMAD2 in myoepithelial cell lines generated as in (a), and stimulated with 5 ng/mL TGFβ (time in min). HSC70 serves as loading control. Images are representative of three experiments. e Invasion assay for MDA-MB-231 (231) and MCF-7 cells using conditioned media (CM) isolated from myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 3 biological replicates, 4-6 technical replicates; ****P < 0.0001 (two-tailed t-test). f Array analysis for proteases using conditioned media isolated from myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 1 biological replicate, 2 technical replicates. g, h Bar graphs showing qRT-PCR analysis for MMP9 (g) and MMP13 (h) using RNA isolated from myoepithelial cell lines generated as in (a) (error bars, +s.e.m). n = 3 biological replicates, 3 technical replicates; ****P < 0.0001 (two-tailed t-test).
