Figure 7
Epithelial-to-mesenchymal transition gene expression changes were confirmed using cultured cells. Cells used were the same as those xenografted onto the chorioallantoic membrane (see Figures 1 and 2). (A) Target gene validation was confirmed by immunoblotting. For either the transforming growth factor-beta receptor II control (TβRIIfl/fl) cells or the transforming growth factor-beta receptor II knockout (TβRII KO) cells, the conditions were as follows: cells alone, cells treated with 1 ng/ml transforming growth factor beta (TGF-β) for 2.5 hours, cells treated with partial TβRII KO fibroblast conditioned media for 24 hours (two cell lines used), or cells treated with TβRIIfl/fl fibroblast conditioned media for 24 hours. Conditioned media treatment from partial TβRII KO and TβRIIfl/fl fibroblasts gave similar results. (B) Wnt11, Dsc2, and Tmeff1 expression in TβRII KO cells paralleled results seen in array results. For each condition (epithelial cells alone or fibroblast conditioned media treatment), all TβRII KO cell samples were respectively compared with TβRIIfl/fl cells. (C) Expression of Nodal signaling inhibitors (Dact2, SnoN) was downregulated but unaccompanied by significant expression increases of Nodal targets. For each condition (epithelial cells alone or fibroblast conditioned media treatment), all TβRII KO cell samples were respectively compared with TβRIIfl/fl cells. CM, conditioned media; fib, fibroblasts.