Extended Data Fig. 8: Antitumour response of Foxp3CreFasnfl/fl mice.
From: Lipid signalling enforces functional specialization of Treg cells in tumours
a, b, Foxp3CreFasn+/+ or Foxp3CreFasn+/fl control (a, n = 9; b, n = 13) and Foxp3CreFasnfl/fl (a, n = 10; b, n = 16) bone-marrow chimaeras were inoculated with B16 cells and euthanized on day 17 (a) or days 17–19 (b). Shown are relative frequencies of TNFα+ cells (left) and IFNγ+ cells (right) amongst conventional CD8+ T cells (a) and Foxp3–CD4+ T cells (b). c–h, Foxp3CreFasn+/+ or Foxp3CreFasn+/fl control (n = 9) and Foxp3CreFasnfl/fl (n = 10) bone-marrow chimaeras were inoculated with B16 cells and euthanized on day 17. c, d, Numbers of conventional CD8+ T cells (c) and Foxp3–CD4+ T cells (d) in PLNs (left) and tumours (right; normalized to tumour weight). e, f, Frequencies of CD44hiCD62Llo cells amongst conventional CD8+ T cells (e) and Foxp3–CD4+ T cells (f). g, Frequency (left) and number of Foxp3+CD4+ T cells in PLNs (middle) and tumours (right; normalized to tumour weight). h, Ratio of CD8+ T cells to Treg cells. i, Foxp3CreFasn+/+ or Foxp3CreFasn+/fl control (n = 13) and Foxp3CreFasnfl/fl (n = 16) bone-marrow chimaeras were inoculated with B16 cells and euthanized on days 17–19. Shown is the relative frequency of IFNγ+ cells amongst Foxp3+CD4+ Treg cells in PLNs and tumours. j, k, GSEA enrichment plots showing gene signatures of fatty-acid metabolism (j) and genes downregulated in TCR-α-deficient aTreg cells compared with TCR-α-sufficient aTreg cells (k), in intratumoral Treg cells from Foxp3CreFasn+/+ or Foxp3CreFasn+/fl control and Foxp3CreFasnfl/fl mice (n = 3 samples in each group) bearing B16 melanoma. l, Treg cells from Foxp3CreFasn+/fl and Foxp3CreFasnfl/fl mice were used for in vitro suppression assays (n = 3 technical replicates) at multiple ratios of Treg cells to naive CD4+ T cells (TN cells). m, n, Treg cells from Foxp3CreFasn+/fl and Foxp3CreFasnfl/fl mice were labelled with CellTrace Violet (CTV), and cultured with anti-CD3/28 antibodies plus IL-2 for 3 days. Shown are flow-cytometry analysis of the frequency of CTVlo cells (m) and the relative expression of GITR (n, left) and CD44 (n, right) (n = 6 per genotype). o, Relative expression of GITR (left; n = 16 per genotype) and CD44 (right; n = 15 per genotype) on Treg cells in PLNs and tumours of mice bearing B16 melanoma. *P < 0.05; **P < 0.01; two-tailed unpaired Student’s t-test (a–i, l, n, o). Data are means ± s.e.m. in a–i, l, n, o. Data are representative of four (m) or two (l) or compiled from three (b, i) or two (a, c–h, n, o) independent experiments. Values in control samples were set to 1 (a, b, i, n, o).
