Fig. 6: Activated PC is downregulated in IBD and limits thrombogenic CD4+ T cell activity. | Nature Communications

Fig. 6: Activated PC is downregulated in IBD and limits thrombogenic CD4+ T cell activity.

From: Tissue factor-dependent colitogenic CD4+ T cell thrombogenicity is regulated by activated protein C signalling

Fig. 6

a PROC (Protein C; PC) expression in colonic biopsies from patients in the RISK study (GEO ID GSE57945) (iCD, n = 162, cCD, n = 56, UC, n = 62, control non-IBD group, n = 42). b, c Following cell culture, T cells were washed and incubated with fluorescently labelled activated PC. Binding was measured by flow cytometry. Following activated PC pre-treatment, activated PC was removed, θ (unactivated cells) and Th0 cells were washed with EDTA-containing PBS, and their capacity to initiate clotting was analysed by d–g thrombin generation and h clot formation assays. Activated PC pre-treated Th1 cell-dependent thrombin generation was analysed by i–l thrombin generation assays and m clot formation assays. Following activated PC pre-treatment, Th0 procoagulant activity was analysed by n FXa generation assay, o F3 gene expression and r, s PDI cell surface expression by flow cytometry. Similarly, Th1 cell thrombogenicity was analysed by p FXa generation assay, q F3 gene expression and t, u PDI cell surface expression by flow cytometry. The contribution of PDI-mediated TF decryption was assessed by treating Th0 (v, w) and Th1 (x, y) cells with 10 mM rutin for 1 h. Cells were washed with EDTA-containing PBS, and their capacity to initiate clotting was analysed by v–y thrombin generation. Student’s t-test (two-tailed) (c), Mann–Whitney U Test (two-tailed) (a, n–q), Student’s paired t-test (two-tailed) (e–g, j–l, m, s, u, w, y), or Wilcoxon test (h) was used to determine statistical significance. Data is expressed as mean ± s.e.m. (a, c, e–h, j–l, m–q, s, u, w, y) for a 322 biological donors (iCD, n = 162, cCD, n = 56, UC, n = 62, control non-IBD group, n = 42), c 5–10 biological donors (n = 5 θ, n = 10 Th0 and n = 9 Th1), e–g, j–l 6 biological donors, h, m 10 biological donors, n, p, s, u 4 biological donors, o, q 8 biological donors and w, y 3–5 biological donors (n = 3 θ, n = 5 Th0/Rutin and n = 5 Th1/Rutin). Source data are provided in the Source Data file.

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