Fig 1: Characterization of CXCR6- and CX3CR1-expressing cells in proliferative diabetic retinopathy epiretinal fibrovascular membranes. Immunoreactivities for CXCR6 (A) and CX3CR1 (B) were detected in vascular endothelial cells. Immunoreactivity for CXCR6 was also detected in stromal spindle-shaped cells (A) (scale bar, 10 µm).
Fig 2: Characterization of cells expressing transmembrane chemokines and their receptors in proliferative vitreoretinopathy epiretinal fibrocellular membranes. Immunohistochemical stainings for CXCL16 (A), CXCR6 (B), CX3CL1 (C), and CX3CR1 (D) showing immunoreactivities in spindle-shaped myofibroblasts. Double immunohistochemistry for CD45 (brown) and CXCL16 (red) (E) or CX3CL1 (red) (F) showed co-expression in leukocytes (arrows). No counterstain to visualize the cell nuclei was applied in panels E and F (scale bar, 10 µm).
Fig 3: Human retinal microvascular endothelial cells (HRMECs) express CXCL16, CXCR6, CX3CL1, and CX3CR1. HRMECs were left untreated or treated with interleukin-1 beta (IL-1ß) (50 ng/ml) or tumor necrosis factor-alpha (TNF-a) (50 ng/ml) for 24 h. Protein expression of CXCL16 (A), CXCR6 (B), CX3CL1 (C), and CX3CR1 (D) in cell lysates was determined by Western blot analysis (representative Western blots are depicted on top of the graphs). The same loading control (ß-actin) was used for quantitation of the relative band intensity of both CXCL16 and CXCR6. Levels of CX3CL1 were quantified in the culture media by ELISA (E). The box plots (median and interquartile range) show results from three different experiments performed in triplicate. Kruskal-Wallis test and Mann-Whitney tests were used for comparisons between three groups and two groups, respectively. *P < 0.05 compared with values obtained from untreated cells. #p < 0.05 compared with IL-1ß-treated cells.
Fig 4: The proinflammatory cytokine tumor necrosis factor-alpha (TNF-a) induces the expression of CXCL16 and ADAM17 in Müller cells. Müller cells were left untreated or treated with TNF-a (50 ng/ml) for 24 h. Levels of CXCL16 were quantified in the culture media by ELISA. Protein expression of CXCL16, CXCR6, ADAM10, and ADAM17 in cell lysates was determined by Western blot analysis. Results are expressed as median (interquartile range) from three different experiments performed in triplicate. (*p < 0.05; Mann-Whitney test).
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