Extended Data Fig. 6: Circulating ADP-heptose in plasma is sufficient to induce TIFAsome formation in human pre-leukaemic cells.
From: Microbial metabolite drives ageing-related clonal haematopoiesis via ALPK1
a, Standard curve generated by measuring TIFAsome formation by THP1-TIFA-tdTomato-GFP cells when stimulated with increasing concentrations of ADP-heptose from 0–100 µg/mL for 30 min, and evaluated on the ImageStream analyzer. b, Representative TIFAsome formation by THP1-tdTomato cells when stimulated with the plasma of IBD patients. Shown is a representative result of 3 independent replicates. Scale bar, 10 µM. c, TIFAsome formation assay using THP1-TIFA-tdTomato-GFP cells (WT and ALPK1-deficient) represented as percent positive cells following incubation with the indicated human plasma samples (n = 3 from independent biological replicates). d, Correlation of plasma ADP-heptose levels as extrapolated from the TIFAsome formation assay with the age of healthy young (< 65 years, n = 11), old (≥65 years, n = 18), MDS (n = 29), CHIP (n = 59), and IBD (n = 8) patients. e, Risk assessment of CHIP individuals with positive (n = 24) or negative (n = 35) ADP-heptose in circulation. Error bars represent the SEM. *, P < 0.05, **, P < 0.01, ***, P < 0.001. P values were calculated for the indicated comparisons using a two-tailed unpaired Student’s t-test (d and e) and a two-way ANOVA (c). FACS-gating schemes are shown in Supplementary Fig. 2.
