Fig 2: Characterization of LPF subsets in OCT in TAO(A) Volcano plot showing DEGs of LPF subsets in TAO/HC comparison.(B) Venn diagram showing comparative analysis of upregulated DEGs of LPF subsets in three fibroblast subtypes and upregulated DEGs of LPF subsets in TAO/HC comparison. The count shows the number of DEGs.(C) Heatmap of the relative expression of selected LPF-specific upregulated DEGs in HC and TAO groups in different fibroblast subtypes.(D) Bar chart showing representative GO terms and KEGG pathways enriched in upregulated DEGs of LPF subtypes in TAO/HC comparison.(E) UMAP plot showing the expression of ACKR1 and CXCR2 of OCT cells from HCs and individuals with TAO.(F) Pseudotime-ordered single-cell expression trajectories for FABP4, FABP5, CEBPB, CEBPD, PLIN2, PPARG, RASD1, and PIM1.(G) Representative images of immunostaining of cross-sections of OCTs from HCs and individuals with TAO for PIM1 (green) and nuclei (4',6-diamidino-2-phenylindole [DAPI]; blue).Each group contains six samples. Data are represented as mean ± SEM. Significance was determined using unpaired Student’s t test. *p < 0.05.

Fig 3: Inhibiting PIM1 alleviated EAU.a Immunostaining of cross-sections of CDLNs from day 0 group and day 14 EAU group shows PIM1 (green) and nuclei (4',6-diamidino-2-phenylindole [DAPI]; blue). Each group contains six mice. P(EAU-Normal) = 0.0003. Data represented as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. ***P < 0.001. Scale bars, 20 mm. b Representative fundus images and clinical scores of eyes from the vehicle group and SMI-4a group after immunization at day 14. White arrowheads indicate inflammatory exudation and vascular deformation. Each group contains six mice. P(EAU-SMI-4a) = 2.8E-06. Data represented as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. ****P < 0.0001. c Representative histopathological images (hematoxylin and eosin staining) and pathological scores of eyes from the vehicle group and SMI-4a group after immunization at day 14. Black arrowheads indicate infiltration of inflammatory cells and retinal folding. Each group contains six mice. P(EAU-SMI-4a) = 1.0E-08. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. ****P < 0.0001. Scale bars, 20 mm. d–f Proportions of Th1 cells (d), Th17 cells (e) and regulatory T cells (Treg) (f) were measured by flow cytometry after immunization at day 14. Each group contains six mice. P(Th1) = 0.0005, P(Th7) = 0.0014, P(Treg) = 3.8E-06. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. *P < 0.05, **P < 0.01, ****P < 0.0001. g–i Proportions of PIM+ cells in Th1 cells (g), Th17 cells (h) and regulatory T cells (Treg) (i) were measured by flow cytometry after immunization at day 14. Each group contains six mice. P(Th1) = 1.8E-07, P(Th7) = 7.4E-08, P(Treg) = 3.5E-07. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. ****P < 0.0001.

Fig 4: Upregulation of PIM1 in VKH patients.a t-SNE plots of T cell subsets from all healthy controls and VKH samples. b t-SNE plots of B cells subsets from all healthy controls and VKH samples. c Violin plots of PIM1 expression in T cell subsets of healthy controls and VKH patients. d Violin plots of PIM1 expression in B cell subsets of healthy controls and VKH patients. e Heatmap of the average expression of PIM1 by PCs and CD4 T cell subsets in healthy controls and VKH patients. f, g The percentage of PIM1+ cells in CD4 T cells (f) and PCs (g) measured by flow cytometry. Each group contains 15 samples. P(CD4 T cells, VKH-HC) = 0.0010, P(PCs, VKH-HC) = 0.0011. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. **P < 0.01, ***P < 0.001. h, i The proliferating rate of CD4 T cells (gated on CD3+CD8- T cells) (h) and B cells (i) measured by flow cytometry. P(CD4 T cells, SMI-4a (-)-SMI-4a (+)) = 6.4E-08, P(PCs, SMI-4a (-)-SMI-4a (+)) = 7.1E-08. Data represented as mean ± SEM from six independent experiments. Significance was determined using unpaired two-tailed student’s t test. ****P < 0.0001.

Fig 5: scRNA-seq analysis of the dynamic changes in T cell subsets.a t-SNE plots of T cell subsets from all mice groups. b Heatmap showing scaled expression of discriminative gene sets for T cell subsets from all mice groups. c t-SNE plots of canonical markers for T cell subsets from all mice groups. d Line charts showing the proportion of T cell subsets in total T cells from EAU and control groups at different time points. e Line charts showing the mean expression of Pim1 in T cell subsets from EAU and control groups at different time points. f Heatmap showing representative GO terms and KEGG pathways enriched in upregulated DEGs of the T cell subsets in the D14c/D0 comparison groups at different time points. Significance was calculated based on the accumulative hypergeometric distribution by Metascape webtool. g Heatmap showing representative GO terms and KEGG pathways enriched in upregulated DEGs of the T cell subsets in the D14/D14c comparison groups at different time points. Significance was calculated based on the accumulative hypergeometric distribution by Metascape webtool.