Immunohistochemistry
The positive IF staining for cytokeratin confirms the epithelial origin
of the cells (figure 2a). In general, protamine treatment increases
extracellular margins between adjacent cells and adherence junction
formation is more disrupted (figure 2d-f). IF analyses showed a clear
signal of occludin all along the cell borders in the controls (figure
2g). In both intervention groups this signal was reduced, and tight
junctions were more interrupted across the cell borders, but still
clearly visible. CS56 shows chondroitin sulfate is present in controls,
but decreases after protamine treatment (figure 2j-k). Even after
removing CS treatment by rinsing the inserts, IF analyses demonstrated
an increased signal in the CS treated samples compared to controls and
protamine alone samples (figure 2l).