Extended Data Fig. 7: TRAP1 deficiency reprograms glutamine and α-KG metabolism in IL-4-stimulated macrophages.
From: Cancer suppresses mitochondrial chaperone activity in macrophages to drive immune evasion
(a) GSEA of glutamine metabolism in Trap1+/+ or Trap1−/− BMDMs stimulated with IL-4 (n = 2). GOBP, Gene Ontology Biological Process. HPO, Human Pathway Ontology. (b, c) Quantification of glutamine consumption (n = 6) (b) and intracellular α-KG (n = 5) (c) in IL-4-stimulated Trap1+/+ or Trap1−/−BMDMs. Data from three independent experiments. (d, e) Activity of SDH (d; n = 3) and αKGDH (e; n = 3) in TAMs from Trap1wt and Trap1cKO tumor-bearing mice. AU, arbitrary units. Data from two independent experiments. (f-h) SDH activity (n = 3) (f), intracellular succinate (n = 4) (g) and α-KG/succinate ratio (n = 4) (h) of IL-4-stimulated Trap1+/+ or Trap1−/− BMDMs. Data are representative of two independent experiments. (i) Schematic of SDH-mediated succinate oxidation and GLS-mediated glutaminolysis. (j) Basal OCR of Trap1+/+ or Trap1−/− BMDMs treated with IL-4 in the presence or absence of DMM or BPTES (n = 6). Data are representative of two independent experiments. (k) α-KG levels in TCM-treated Trap1+/+ or Trap1−/− BMDMs with or without DMM for 24 h (n = 3). Data are representative of two independent experiments. (l, m) Expression of CD206 and CD301 (l), and PD-L2 and RELMα (m) in Trap1+/+ or Trap1−/− BMDMs stimulated with IL-4 in the presence or absence of DMM, measured by flow cytometry (n = 4). Data are representative of three independent experiments. (n) Gene expression in Trap1+/+ or Trap1−/− BMDMs stimulated with IL-4 in the presence or absence of BPTES for 6 h, assessed by RT-qPCR (n = 4). Data are representative of two independent experiments. (o) α-KG levels in TCM-treated Trap1+/+ or Trap1−/− BMDMs with or without diethyl succinate (Suc) for 24 h (n = 3). Data are representative of two independent experiments. (p) Gene expression in Trap1+/+ or Trap1−/−BMDMs stimulated with IL-4 under conditions with or without Suc for 6 h, assessed by RT-qPCR (n = 4). Data are representative of two independent experiments. All data are mean ± s.e.m. and were analyzed using a two-tailed, unpaired Student’s t-test (b-h) or one-way ANOVA with Sidak’s multiple comparison test (j-p).
