Fig. 2. Role of N-cadherin in rTGF-ß1-stimulated invasion of myofibroblasts. (A) 4x105 single myofibroblasts were seeded upon a Matrigel coated filter with rTGF-ß1 or HCT-8/E11 cells in the lower compartment to stimulate invasion. N-cadherin was neutralized by the GC-4 mAb which was added together with the myofibroblasts. Bars indicate means of three results ± s.d. Asterisks show statistically significant inhibition of invasion. (B) Myofibroblast spheroids cultured in collagen type I for 48 hours in the presence of HCT-8/E11 cells, and cultured in collagen type I for 7 days untreated or treated with rTGF-ß1, were challenged with control IgG1 (80 µg/ml) or GC-4 mAb (80 µg/ml). In the 7-day experiment, medium was exchanged on day 3 and 5. A representative phase-contrast micrograph is shown for each condition. Scale bar, 100 µm. (C) Western blot, showing the effect of combinations of siN-CAD oligonucleotides on N-cadherin expression and cadherin-11 expression in myofibroblasts. Tubulin was used as loading control. (D) Spheroid myofibroblasts of cells that had been electroporated with siN-CAD oligonucleotides and cultured in collagen type I for 2 or 4 days in the presence of rTGF-ß1. con, control oligonucleotide. Scale bar, 100 µm.