Vitreous concentrations of angiopoietins in patients newly diagnosed with retinal diseases and cell model of barrier breakdown testing the interaction.

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Vitreous concentrations of angiopoietins in patients newly diagnosed with retinal diseases and cell model of barrier breakdown testing the interaction of VEGF‐A and angiopoietins Vitreous concentrations of angiopoietins in patients newly diagnosed with retinal diseases and cell model of barrier breakdown testing the interaction of VEGF‐A and angiopoietins A, BBox plots of vitreal ANG‐1 (A) and ANG‐2 (B) levels from newly diagnosed patients with wAMD, DR, proliferative DR and RVO compared to controls (macular hole). The interquartile range of the data is indicated by the box. A nonparametric Kruskal–Wallis analysis followed by Dunn's method for multiple comparisons was used to show significant differences of the groups to control which are indicated by asterisks. ANG‐1 levels did not differ significantly, but ANG‐2 levels were significantly different: control vs. AMD (*, P = 0.0451), vs. DR (****, P < 0.0001), pDR (****, P < 0.0001), and RVO (****, P < 0.0001). CSchematic of cellular experiments to measure transendothelial resistance in human endothelial cells using transwell filters and CellZscope technology.D24‐h concentration of ANG‐2 in supernatants of the basal side of the culture stimulated with 5 ng/ml of VEGF‐A, 200 ng/ml of ANG‐1, or the combination. Error bars show SEM with one‐sided ANOVA (P < 0.0001) and Tukey's multiple t‐test for five independent experiments indicating significance for control vs. VEGF‐A (**, P = 0.0028); VEGF‐A vs. ANG‐1 (****, P < 0.0001); ANG‐1 vs. VEGF‐A and ANG1 (**, P = 0.0061); VEGF‐A vs. VEGF‐A and ANG‐1 (ns, P = 0.0625).EHuman endothelial cells plated on filters were assessed for endothelial barrier function over time after the addition of 5 ng/ml VEGF‐A alone or in combination with 200 ng/ml ANG‐1 or 10 μg/ml anti‐ANG‐2 or all three test items. The final time point was used for statistical analysis. Error bars show SEM with one‐sided ANOVA (P < 0.0001) and Tukey's multiple t‐test for three independent experiments indicating significance of VEGF‐A vs. untreated (****, P < 0.0001); vs. VEGF‐A and ANG‐1 and anti‐ANG‐2 (***, P = 0.0004); vs. VEGF‐A and anti‐ANG‐2 (*, P = 0.038); vs. VEGF‐A and ANG‐1 (*, P = 0.040). Furthermore, VEGF‐A and anti‐ANG‐2 and ANG‐1 are significantly different vs. VEGF‐A and ANG‐1 (*, P = 0.0451); vs. VEGF‐A and anti‐ANG‐2 (*, P = 0.0474); vs. untreated (*, P = 0.0492). Finally untreated is significantly different vs. VEGF‐A and ANG‐1 (***, P = 0.0009) and VEGF‐A and anti‐ANG‐2 (***, P = 0.0008).Data information: ANOVA, analysis of variance; SEM, standard error of the mean; ANG‐2, angiopoietin‐2; DR, diabetic retinopathy; RVO, retinal vein occlusion; wAMD, wet age‐related macular degeneration; pDR, proliferative diabetic retinopathy; vs., versus. Jörg T Regula et al. EMBO Mol Med. 2016;8:1265-1288 © as stated in the article, figure or figure legend