Extended Data Fig. 10: Effects of ADP-heptose on gene expression, inflammatory cytokines, and signalling pathways.

a, Pathway enrichment of GSEA and Hallmark datasets in Dnmt3a^WT cells treated with ADP-heptose versus H₂O (left panels) and Dnmt3a^KO cells treated with ADP-heptose versus H₂O (right panels) (n = 3 mice per group). NES, normalized enrichment score. Absolute enrichment score (ES) and corresponding P value are shown for each pathway. b, Representative flow cytometric profile of EdU+ HSCs after ADP-heptose or H₂O treatment. c, Serial colony replating of ADP-heptose- or H₂O-treated BM HSPCs isolated from Dnmt3a^WT or Dnmt3a^R878H mice (n = 3 mice per group). In each plating, colonies were scored at day 14. d, Heatmap of differentially expressed cytokines and chemokines as measured in the BM fluid of Dnmt3a^WT, Dnmt3a^KO, and Dnmt3a^KO;Alpk1^KO mice treated with ADP-heptose (0.5 mg/kg) in vivo (Data are representative of 4 independent biological replicates; for each replicate, BM was pooled from 2 different mice). e, Immunoblotting performed on THP1 cells untreated or treated with ADP-heptose (1 µg/ml) in combination with DMSO (-), UBE2N inhibitor at 5 µM (+), UBE2N inhibitor at 10 µM (+ +), IRAK1/4 inhibitor at 0.25 µM (+), and IRAK1/4 inhibitor at 0.5 µM (++) for 30 min. f, Immunoblot analysis of NF-κB modulators on wildtype (WT), IRAK1^KO, IRAK4^KO, IRAK1/4^dKO, MyD88^KO, and TRAF6^KO THP1 cells upon treatment with ADP-heptose (1 µg/ml) for 30 min. For gel source data, see Supplementary Fig. 1. 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-way ANOVA (c).

Source data