Fig 1: HSP27 antigen-specific mechanisms involved in HIOP-induced glaucoma in mice. HIOP and HSP27-immunized animal models were established as described in Methods. All experiments were performed in triplicate with n = 6 animals per group. A Quantitative analysis of Retinal Ganglion Cells (RGCs) in mouse retinal flat-mounts stained with anti-RBPMS. Representative images of immunofluorescent staining are shown. The images were acquired in black and white to facilitate highly linear quantification of positive cells. Data are presented as means ± SEM. Scale bar = 100 μm. B-D TCR distribution analysis in the retina (B), blood (C), and lymph nodes (D), including total clones, unique clones, and Chao1 index. E Analysis of shared TCRs among the retina, blood, and lymph nodes in HIOP mice. F Venn diagram illustrating shared TCRs distributed in the retina, blood, and lymph nodes. G Analysis of shared TCRs between HIOP retinas and HSP27-immunized T cells. H Sankey diagram of the 12 specific TCRs from HSP27-immunized T cells, visualizing the associations between CDR3 amino acid sequences (CDR3AA) of TCRs and their corresponding variable (V) and joining (J) genes. The three CDR3AA-V-J sequences consistent with retina-specific TCRs in HIOP mice are labeled in red. Differences between groups were compared using One-Way ANOVA. P-values are indicated on the graphs (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)