Figure 5

Mesotrypsin enhances adherens junctions (AJs). (A) Assessment of E-cadherin mRNA (left, n = 3) and protein (right, n = 3, **p < 0.01) expression. In addition, HaCaT cells expressing Venus-mesotrypsin exhibited upregulated E-cadherin. (B) Upper left, A schematic representation of the cell aggregation assay designed to evaluate cadherin activity. Cells subjected to trypsin treatment in the presence of calcium (TC treatment) eliminate most cell adhesion molecules, leaving cadherins intact. Disrupted cells in rotational culture reassembled into aggregates, the size of which reflects the activity of E-cadherin. Middle and lower, Representative images of reaggregated cells. Scale bars, 200 µm. Right, Quantification of aggregate sizes. n = 3, *p < 0.05. The expression of Venus-mesotrypsin, as opposed to Venus alone, results in the formation of large aggregates, indicating heightened cadherin activity.