Fig. 7

Retinal endothelial expression of VCAM-1 is critical for Th1 cell migration across the blood-retina barrier (BRB). A Shown are results representing one of three independent experiments. The mRNA expression of VCAM-1 in the retina of GL mice (day 20) was determined by qRT-PCR. Ctrl, n = 8; GL, n = 6. n refers to the number of RNA tests. Two retinas from 2 mice (one retina from each mouse) were combined for each RNA test. B Shown are results representing one of three independent experiments. Representative confocal images of retina flat-mounts stained for VCAM-1 from Ctr (n = 4) and GL (day 20, n = 4) (scale bar: 500 μm). Fluorescence intensity (FI) of VCAM-1 was quantified. n refers to the number of retinas used for staining and one retina was analyzed per mouse. C Experimental design: glaucoma was induced and mice were intraperitoneally injected with anti-VCAM-1 (αVCAM-1) or isotype antibody every 3 days from day 15 to the end of indicated experiments. D–F Shown are results representing one of three independent experiments. In each experiment, one random retina from each recipient mouse was used for RGC density and the other was for Iba1 and GFAP co-staining (isotype, n = 5; αVCAM-1, n = 4. n refers to the number of retinas). D RGC number, E the percentage of GFAP⁺ area per microscopic field (size: 319.45 μm²), and F the number of microglia (magnification: morphological changes) were quantified. G Shown are results representing one of three independent experiments. Immunofluorescence staining for Isolectin, CXCR3, and DAPI (blue) in retina flat-mounts of GL mice with CD4-specific expression of Tomato, treated with αVCAM-1 or isotype, and retinal CXCR3⁺ CD4⁺ T cells were quantified (isotype, n = 4; αVCAM-1, n = 4. n refers to the number of retinas used for staining and one random retina per mouse was used). D Scale bar: 500 μm; inset, 20 μm. E, F Scale bar: 20 μm. g Scale bar: 20 μm; inset, 5 μm. *P < 0.05, **P < 0.01, ***P < 0.001. two-tailed unpaired Student’s t test was performed