Fig. 3.

Polarized primary human fetal RPE (hfRPE) cell culture model. Cells were exposed to 4-hydroxy-2-nonenal (4-HNE) for 24 h. (a; upper) Phase-contrast microphotographs and microphotographs of the immunofluorescence (IF) staining of hfRPE cultures. (a; lower) The expression of RPE65, a RPE-specific protein, was confirmed through Western blot analysis of the lysate of hfRPE cells. (b) Quantification of transepithelial resistance (TER). (c) Expression of the epithelial marker E-cadherin and the mesenchymal marker vimentin in hfRPE cells exposed to 4-HNE. (d) Immunostaining of drusen-related proteins, APOA1, cathepsin D, clusterin and CFH in hfRPE cells. (e) Western blot analysis showing increased expression of APOA1, cathepsin D, and clusterin in 100 μM 4-HNE-treated hfRPE cells compared to that in controls