Fig. 4

Block of Th1 response ameliorates glaucomatous retinal degeneration. Flow cytometry was performed to determine circulating A Freq. CXCR3⁺ CD4⁺ T cells (Ctr, n = 8; GL, day 20, n = 8) and B Freq. IFN-g⁺ CD4⁺ T cells (Ctr, n = 5; GL, day 20, n = 5). A, B Shown are results representing one of three independent experiments and n refers to the number of used mice. C, D Peripheral blood CD4⁺ T cells from GL_20d (CD4_GL, n = 24) and Ctr mice (CD4_Ctr, n = 24) were obtained by fluorescence-activated cell sorting (FACS). RNA-seq was performed to analyze the gene expression profile in CD4_GL and CD4_Ctr. C Analysis of Gene Ontology (GO) enriched pathways showing significantly altered Th1-associated biological processes (BP) in CD4_GL compared to those in CD4_Ctr. D Heatmap of differentially expressed genes (DEG) involved in Th1 differentiation. E Experimental design: glaucoma was induced and mice were intraperitoneally injected with anti-IFN-g (aIFN-g) or isotype antibody every 3 days from day 15 after MB injection to the end of indicated experiments. F, H 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 staining. n = 5, n refers to the number of retinas for analysis. The number of F RGC, G microglia (magnification: morphological changes), and H the percentage of GFAP⁺ area per microscopic field (size: 319.45 μm²) was calculated. F scale bar: 500 μm; inset, 20 μm. G-H scale bar: 20 μm. **P < 0.01, two-tailed unpaired Student’s t test was performed