Fig. 4: Pim1 regulated the balance of Th17/Treg cell.

a–c CDLN cells from EAU mice were cultured with IRBP1-20 or IRBP1-20 plus SMI-4a (20 μM). Flow cytometry was performed to show the proportion of Th1 cells (a), Th17 cells (b) and Treg cells (c). Each group contains six mice. P(Th1) = 9.3E-09, P(Th7) = 0.0030, P(Treg) = 0.0040. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. **P < 0.01. d, e Representative fundus images (d) and clinical score (e) after induction of CD4⁺ T cells cultured with IRBP1-20 or IRBP1-20 plus SMI-4a at day 14. White arrowheads indicate inflammatory exudation and vascular deformation. Each group contains six mice. P(EAU-SMI-4a) = 3.8E-05. Data expressed as mean ± SEM. Significance was determined using unpaired two-tailed student’s t test. ****P < 0.0001. f, g Representative fundus images (f) and clinical score (g) after induction of Pim1 shRNA treated-CD4⁺ T cells cultured with IRBP1-20 at day 14. Each group contains six mice. P(CD4-CD4 + Nc shRNA) = 0.9552, P(CD4-CD4 + pim1 shRNA) = 0.0002, P(CD4 + Nc shRNA- CD4 + pim1 shRNA) = 0.0003. Data expressed as mean ± SEM. Significance was determined using one-way ANOVA. ns no significant differences, ***P < 0.001. h–j CD4 T cells from EAU group cultured with IRBP1-20 alone or with IRBP1-20 plus with SMI-4a for 72 h. Flow cytometry showed the proportion of PIM1⁺ cells (h), pAKT⁺ cells (i), and pFOXO1⁺ cells (j) in total CD4⁺-gated T cells. Data represented as mean ± SEM from six independent experiments. P(PIM1⁺ cells,Control-IRBP1-20) = 0.0208, P(PIM1⁺ cells, IRBP1-20-IRBP1-20+SMI-4a) = 8.2E-09, P(pAKT⁺ cells,Control-IRBP1-20) = 3.8E-12, P(pAKT⁺ cells, IRBP1-20-IRBP1-20+SMI-4a) = 8.6E-10, P(pFOXO1⁺ cells cells,Control-IRBP1-20) = 4.4E-07, P(pFOXO1⁺ cells, IRBP1-20-IRBP1-20+SMI-4a) = 1.8E-06. Significance was determined using two-way ANOVA. **P < 0.01, ****P < 0.0001.