Fig. 1

Chronic exposure to activated monocyte supernatants or TNFα increases the percentage of enlarged, multinucleated RPE cells. Porcine retinal pigment epithelium (RPE) cells were cultured with or without lipopolysaccharide activated monocyte supernatants (Mo SN) or TNFα added daily at different concentrations for a total of 10 days. a Representative pictures of Zonula Occludens (ZO)1 (green) and Hoechst (blue) immunohistochemistry of control, Mo SN, and TNFα-exposed RPE. Automated quantifications by Arrayscan of b the number of RPE cells, c RPE cell size, d the number of multinucleated RPE cells (≥ 2 nuclei), and e total number of nuclei per mm² in the indicated different conditions. Representative images of 10-day TNFα-exposed RPE cell-cultures of f a co-culture of 50% RPE cells labeled with Carboxyfluorescein succinimidyl ester (CFSE) (green) and 50% RPE cells labeled with Far-red CFSE (red), and g ZO-1- (green) and Hoechst nuclear- (blue) stain of cultures maintained in the presence of EDU (5-ethynyl-2′-deoxyuridine, red). (n = 5/group, one way ANOVA or Mann-Whitney, comparison versus control: *P < 0.05, **P < 0.005*** P < 0.0005)