E-cadherin loss contributes to reduced substrate adhesion in MCF10A
CDH1-/-. Both isogenic cell lines were seeded at a density of 2.0 × 104 cells and images were taken at 2 h intervals for up to 8 h using the IncuCyte 2011A FLR. a) Representative images of cell substrate adherence at 4 h post seeding under uncoated conditions. b) Histogram representing the number of adhered to non-adhered cells from both cell lines under uncoated conditions over different time points. Significantly reduced substrate adhesion were observed in MCF10A CDH1-/- cells as determined by student t-test p < 0.01 (represented by **). c) Histogram representing the number of adhered to non-adhered cells from both cell lines under different ECM coating conditions (UC, uncoated; COL, collagen; LN, laminin; FN, fibronectin; VN, vitronectin) at 4 h post seeding. For each condition, three images were acquired and the population of non-adhered cells to total cell numbers from each image were counted by two independent researchers and the average taken. Results of experiments done under different ECM coatings showed MCF10A CDH1-/- cells consistently showed reduced substrate adhesion compared to the wildtype cells (student t-test; p < 0.01 represented by **). Furthermore, different ECM coatings provided varying increases in the numbers of adhered cells in both isogenic cells.