Fig. 7: 2-KG decreases ammonia and restores survival of GOT1-deficient human CD8+ T cells.
From: The malate shuttle detoxifies ammonia in exhausted T cells by producing 2-ketoglutarate
a, Experimental design of b. Healthy donor CD8+ T cells were electroporated with Cas9 protein and GOT1-specific gRNA and were differentiated into Tex-like cells or Tmem-like cells. b, Western blot images show the expression of GOT1 in the indicated populations of cells. GRP94 was used as a loading control. A bar graph shows the results of densitometric quantification of the GOT1 immunoblot bands. c, Illustration of the experimental design of d and e. d, Bar graph shows the concentrations of ammonia in CD8+ T cells. e, A line graph shows the numbers of cells on day 1 and day 2 after restimulation with anti-CD2, anti-CD3 and anti-CD28. Data are cumulative from two independent experiments with eight healthy donors in total. Results are mean ± s.d. ***P < 0.001; ****P < 0.0001. Comparisons were performed with one-way ANOVA (b,d) or two-way ANOVA (e). In b, P = 0.000192726 and P < 1.0 × 10−15; in d, both P < 1.0 × 10−15; in e (from top to bottom), P = 6.4 × 10−11 and P = 6.7 × 10−8.
