Extended Data Fig. 1: TRAP1 expression negatively correlates with immunosuppression in human and murine tumors.
From: Cancer suppresses mitochondrial chaperone activity in macrophages to drive immune evasion
(a) Representative gating strategy used for flow cytometry analysis of immune cells. (b) Heatmap showing TRAP1 expression in macrophages, monocytes, neutrophils (Neu), DC, NK, CD8+ T and CD4+ T cells isolated from the brain, liver, peritoneal cavity (PEC), lymph nodes (LN), spleen (SP), blood, and bone marrow (BM) tissues of naïve wild-type mice (n = 5 mice per group). Data are representative of three independent experiments. (c) Frequencies of TRAP1+ macrophages (Mac), monocytes (Mono), neutrophils (Neu), DC, NK, CD8+ T and CD4+ T cells in tumors from YUMM1.7 tumor-bearing mice on day 16 (n = 5 mice per group). Data are representative of two independent experiments. (d) Immunoblot analysis for TRAP1, VDAC1/2, prohibitin 1 (PHB1), GAPDH, and α-Tubulin in the mitochondrial and cytosolic fractions of naïve wild-type macrophages. Data are representative of two independent experiments. (e) Single-cell RNA-seq dataset (GSE146771) analysis of CD86 and CD163 expression in TRAP1hi and TRAP1lo macrophages from patients with colon cancer. (f) Pearson correlation of TRAP1 expression with genes encoding molecules associated with immune response in patients with skin cutaneous melanoma from the TCGA. (g) Representative flow cytometry plots (left) and quantitative analysis (right) of TRAP1+ macrophages in the spleen and tumor from B16-F10 tumor-bearing mice on day 16 (n = 5 mice per group). Data are representative of two independent experiments. (h, i) Quantitative analysis of TRAP1+ macrophages (h) and CD206+ macrophages (i) in the peritoneum, spleen, and tumor of YUMM1.7 tumor-bearing mice on day 16 (n = 5 mice per group). Data are representative of two independent experiments. (j) Expression of indicated genes in BMDMs treated without (Ctrl) or with YUMM1.7 TCM for 24 h, assessed by RT-qPCR analysis (n = 4). Data are representative of four independent experiments. All data are mean ± s.e.m. and were analyzed using a two-tailed, paired Student’s t-test (g), unpaired t-test (e), or a one-way ANOVA with Sidak’s multiple comparison test (h, i).
