Fig. 3: SNX9 KO improves effector functions of exhausted T cells and dampens the NFAT-NR4A1/3-TOX axis.
From: Deletion of SNX9 alleviates CD8 T cell exhaustion for effective cellular cancer immunotherapy
a Schematic procedure to generate SNX9 KO T cells. b IFNγ secretion by ELISA of Teff and Tex in response to re-stimulation. n = 10 donors (Teff) and 8 (Tex). c Top row: representative confocal images of Tex co-cultured with T2 tumor cells for actin (n = 6), and LAMP1/Perforin (n = 3). Bottom row: example images of CD28-EGFP (n = 6) or TCRz-EGFP (n = 5) fusion proteins, or anti-CD45 (n = 6) and SNX9 in Teff. All images are provided in the Source Data file. d Nuclear translocation measured by image cytometry of NFATc2 in Teff upon anti-CD3/28 stimulation. n = 7 donors of n = 2 experiments. e Area under the curve (AUC) of Calbryte520-AM Ca2+ flux upon stimulation. n = 8 donor of n = 3 experiments. f RT-qPCR quantifications of mRNA for NR4A1 and NR4A3 in cells on day 6 of the Tex culture. n = 7 donors of n = 2 experiments. g Antibody staining (delta unstained) of TOX in Tex. n = 8 donors of n = 4 experiments. h Expression of CCR7 measured by flow cytometry antibody staining in Tex. n = 8 of n = 3 experiments. b, e-h Statistics are a two-sided paired t-tests. i Geometric mean CD25 upregulation relative to unstimulated cells for Teff co-cultured with T2 wt or T2 CD80 CD86 KO (T2 KO) cells. 10 μg/ml human IgG1 or 10 μg/ml anti-CTLA4 blocking antibody (ipilimumab) was added. n = 6 donor replicates of n = 2 experiments. j CD25 geometric mean intensity of stimulated T cells with or without SNX9 KO. n = 7 donors of n = 2 experiments. k Delta unstimulated geometric mean of phospho-PLCγ1-Tyr783 fluorescence intensity. n = 4 donors. d, i-k Statistics are paired 2-way ANOVA with Holm-Sidak correction. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. Source data and exact p-values are provided as a Source Data file.
