Fig 2: ITK inhibitor soquelitinib mitigates exhaustion in CD19 CAR-T cells in vitro.A–D Summary data showing the percentages of PD-1+, TIM3+, TIGIT+, and LAG3+ cells within CD4+ and CD8+ CD19 CAR-T cell populations after 7-day exposure to soquelitinib (0-40 nM) with anti-CD3/CD28 bead activation and IL-2 (500 IU/mL); n = 4–6. E–H Summary data showing the percentages of LAG3+, TIGIT+, TIM3+, and CD39+ cells within CD4+ and CD8+ CD19 CAR-T cell populations after 7-day exposure to soquelitinib (0–40 nM). CD19 CAR-T cells were co-cultured with HBL-1 tumor cells and subjected to repetitive antigen stimulation over a 7-day period; n = 3. I, J Summary of percentages of 2B4+, CD39+ and CD38+ cells within CD4+ and CD8+ CD19 CAR-T cell populations; n = 4–5. K, L The expression of nuclear transcription factors BATF was analyzed within CD4+ and CD8+ CAR-T cells by flow cytometry; n = 5. M The expression of p-PLCγ-1and PLCγ-1 were analyzed by western blotting (GAPDH used as loading control). N The expression of transcription factors TCF1, NR4A, NFAT1, TOX and BATF were analyzed by western blotting (GAPDH used as loading control). O The expression of IFN-γ, TNF-α and LAG3 were analyzed by western blotting (GAPDH used as loading control).

Fig 3: Increased TOX levels in retinal endothelial cells (ECs) in Cdh5CreER;Tgfbr1CKO/- mice.(A) Uniform Manifold Approximation and Projection (UMAP) plots for Tox and Tox3, two members of the Tox transcription factor (TF) family that show increases in transcript abundance in Cdh5CreER;Tgfbr1CKO/- CNS ECs. Red arrows, EC cluster. UMAP plots are similar to those in Figure 9C. (B) Retina flatmounts immunostained for TOX and PECAM1. Arrows indicate TOX+ EC nuclei. The TOX signal is several-fold more intense in the EC nuclei in Cdh5CreER;Tgfbr1CKO/- retinas compared to control retinas. For each genotype, three retinal depths are shown: nerve fiber layer (left), ganglion cell layer (center), and inner plexiform layer (right). Many non-ECs express TOX.