Fig. 6.

TNFα-induced TGF-ß expression mediates the RPE polynucleation and impairment of barrier properties. a TGFβ 1-mRNA expression normalized with S26 expression quantified by RT-qPCR of 2, and 10-day RPE culture exposed to 20 ng/ml of TNFα. b Representative pictures of Zonula Occludens (ZO)1 (green) and Hoechst (blue) immunohistochemistry of control, TNFα(20 ng/ml), and TNFα(20 ng/ml) + antiTGFβ (500 nM)-exposed RPE. Quantifications of the number of multinucleated RPE cells (≥ 2 nuclei) in the indicated conditions. (a, b, n = 5/group, one-way ANOVA or Mann-Whitney versus control * P < 0.05, **P < 0.005, ***P < 0.0005). c Representative pictures of RPE staining with Zonula occludens (ZO) 1 (green) and F actin (Phalloidin, red) after 10 days of culture in the indicated conditions. d RPE transepithelial electric resistance (TER) measured daily in a 10-day trans-well culture with or without TNFα (20 ng/ml) and TNFα (20 ng/ml) + antiTGF-β (500 nM) (n = 3/group, *P < 0.05 one-way ANOVA of control versus TNFα and TNFα + antiTGF-β for each time point, ϒ P < 0.05 Mann-Whitney of TNFα + antiTGF-β versus TNFα for each time point)