Extended Data Fig. 7: IRE1α–XBP1 signalling alters function of T cells associated with ovarian cancer, and promotes malignant progression.

a, Transcriptional profiling of splenic CD44^highCD62L^lowCD4⁺ T cells sorted from naive wild-type versus XBP1-deficient mice. Expression of the differentially regulated genes identified in Fig. 4a is shown (n = 3 per group). b, c, e, f, Analysis of wild-type versus XBP1-deficient CD4⁺ T cells isolated from mice that bore metastatic ovarian cancer (n = 4 per group). b, Expression of previously reported target genes of regulated IRE1α-dependent mRNA decay (RIDD). c, Relative gene expression of master transcription factors controlling helper T cell differentiation. d, Intracellular staining for FoxP3 (left) and proportion of FoxP3⁺CD4⁺ T cells from wild-type (n = 5) and XBP1-deficient (n = 6) mice that bore metastatic ovarian cancer for 21 days. e, Predicted upstream regulators associated with the transcriptional changes observed. f, Enriched cellular functions in XBP1-deficient CD4⁺ T cells at tumour sites. Z-scores greater than 2 indicate functions predicted to be significantly increased in XBP1-deficient CD4⁺ T cells. g, Intracellular staining for IFNγ in CD45⁺CD3⁺CD4⁺ T cells from wild-type and conditional XBP1-deficient mice that bore metastatic ovarian cancer for 29 days (late stage). Representative plots from three independent experiments. h, Intracellular staining for perforin in CD45⁺CD3⁺CD8⁺ T cells from wild-type and conditional XBP1-deficient mice that bore metastatic ovarian cancer for 23 days. i, Intracellular staining for IFNγ in CD45⁺CD3⁺CD8⁺ T cells from wild-type and conditional XBP1-deficient mice that bore metastatic ovarian cancer for 29 days (late stage). Representative plots from two independent experiments. j, In vivo glucose uptake by CD44^highCD4⁺ tumour-infiltrating lymphocytes in Xbp1^f/f (n = 6) or Xbp1^f/fCd4^cre (n = 7) female mice that bore metastatic ovarian cancer. k, Representative TMRE staining for CD45⁺TCRβ⁺CD44⁺CD4⁺ T cells associated with ovarian cancer, from tumour-bearing Xbp1^f/f (n = 3) or Xbp1^f/fCd4^cre (n = 4) mice. l, Peritoneal wash samples were collected from mice at 24 days after tumour challenge and the proportion of CD4⁺ T cells associated with ovarian cancer that expressed PD-1, CTLA4 and TIM3 in wild-type (n = 9) or XBP1-deficient (n = 8) mice was determined. m, Female mice (n = 4 per group) were implanted with ID8-Defb29/Vegfa ovarian cancer cells in the flank, and tumour growth was monitored over time (left). Tumours were resected at day 34 and final size was confirmed ex vivo (right). n, Ascites accumulation (left) in Ern1^f/f (n = 5) or Ern1^f/fCd4^cre (n = 6) mice that bore ID8-Defb29/Vegfa ovarian cancer and overall survival (right, n = 6 per group). Ern1 is the gene that encodes IRE1α. Data are shown as mean ± s.e.m. (c, d, j–l). Boxes represent median ± interquartile range and whiskers indicate minimum and maximum (m, n). n values represent biologically independent mice (a–d, j–n). Two-tailed Student’s t-tests (d, j–l, m, n); log-rank test for survival (n). *P < 0.05, NS, not significant; gMFI, geometric mean fluorescence intensity.

Source data