Figure 4. ICC results for confluent hRPE cultures. Cells were immunostained for Claudin-1 (A, B), N-cadherin (C, D), β-catenin (E, F) and ZO-1 (G, H). I-L: Percentage of intercellular junctions covered by the corresponding protein in cultures of hRPE cells grown for 1 day or 4 weeks after reaching confluency. All junctional proteins, except for ZO-1, had limited localization after 1 day post confluence. In all cases, junctional localization increased markedly after 4 weeks. Data represent the mean ± standard deviation for three replicates from three representative confocal images per each time point for each junctional protein (n = 3). * p<0.05, ** p<0.01. M. Z-Stack scan for a long-term (4 weeks after confluency) culture of hRPE cells grown on porous cell culture inserts, confirming apical localization of ZO-1 and basolateral localization of VEGF. Scale bar = 25 μm.