Figure 4.

Dissociated cells have significantly lower E-Cadherin expression. (a) Histograms of E-Cadherin stain density per unit cell membrane area for HCT-8 cells on 21 kPa substrates on the seventh day of culture. The two histograms are for cells before and after dissociation. Panels a1 and a2 show confocal images of E-Cadherin stain of cells before and after dissociation, respectively. The open arrowheads in panel a1 indicate the continuous and dense E-Cadherin distribution along the cell-cell contact borders. The open arrowheads in panel a2 indicate a weak E-Cadherin staining pattern on the disassociated cells' membrane and the cell-cell contact regions. Panels a1′ and a2′ are the magnified views of the shaded squared areas in panels a1 and a2, respectively. (b) Histograms of E-Cadherin staining density per unit cell membrane area for original HCT-8 cells (not exposed to 21 kPa PA gels) and recultured disassociated HCT-8 cells (that had been cultured on 21 kPa PA gels for 14 days) on stiff polystyrene substrates (3.6 GPa) on the fourth culture day. The open arrowheads in panel b1 indicate the rich E-Cadherin presence along the cell-cell contact borders. The open arrowheads in panel b2 indicate a weak E-Cadherin pattern on the recultured disassociated cells' membrane. Panels b1′ and b2′ are magnified views of the shaded squared areas in panels b1 and b2, respectively. Scale bars: 10 μm. n = 35 for each histogram.