MCF-7s and NBFs form heterotypic adhesions and express cadherin-23 <i>in vitro</i>.

<p>A) NBFs were pre-labeled with CellTracker™ Green and co-cultured with MCF-7s for 3 days. Cells were then fixed and labeled with an antibody against cytokeratin-8 (red). Arrow shows a site of heterotypic contact between the two cell types. B) Three-day co-cultures labeled with antibodies against β-catenin (red) and cytokeratin-8 (green). Arrow – heterotypic contact; Arrowhead – MCF-7 homotypic contact. Scale = 10 µm. C) MCF-7s labeled with CellTracker Green™ (arrowhead) were co-cultured with NBFs labeled with CellTracker Red™ (arrow). Phase contrast (left) and fluorescence image (right) of a typical heterotypic cell aggregate are shown. Scale = 20 µm. D) Immunoblots show no vimentin expression in MCF-7 fraction and no cytokeratin-8 in NBF fraction after FACS separation of co-cultures. PCR (E) and immunoblots (F) showing cadherin-23 expression in MCF-7s and NBFs before and after co-culture. α-tubulin and GAPDH were used as controls.