Figure 4. Comparison of laminin beta 3 and collagen IV expression in different 3D cultures. A: Duplex IHC for laminin beta-3 and collagen type IV in corneal organotypic cultures. The CFib + Epi cultures showed high expression of laminin beta-3 (green) in basal and more superficial epithelial layers. Collagen type IV (red) at the interface of the epithelial cells and fibroblasts indicates a normally regenerated epithelial BM. However, CMyo + Epi cells in organotypic cultures showed no evidence for epithelial BM generation, although myofibroblasts were found to produce collagen type IV. In Epi only controls, laminin beta-3 is expressed throughout the epithelium, with low expression of collagen type IV. Blue is DAPI staining of cell nuclei. Mag.: 200×. B: Graph showing percentage formation of laminin beta-3 and collagen IV compared between the groups of 3D cultures. Error bars represent mean ± SEM. One-way ANOVA was used to compare between the groups, and p < 0.05 was considered statistically significantly different.