Extended Data Fig. 9: Choline supplementation modulates lipid composition and preserves HSPC function.
a, Gating scheme for apoptosis quantification by Annexin V and DAPI staining. b, Apoptosis quantification in mouse HSCs after 48 h choline treatment using Annexin V and DAPI staining. c, Heatmap of all detected lipids passing quality thresholds in untargeted lipidomics of choline-treated and control HSCs. The log2FC values between paired choline-treated and control HSCs per individual are depicted. n = 6. d, Barplots of the summed signal intensity of lipids per class in choline-treated and control HSCs. Subclasses were based on unsaturation levels or acyl-chain length. Paired two-sided Student’s t test or Wilcoxon test. e, Representation of incorporation of 13C-labelled choline in metabolomics and lipidomics after 72 h treatment in human HSPCs. Pie charts depict the ratio of 13C-labelled metabolites or lipid species per class based on unsaturation levels and acyl-chain length. Size in lipid pie charts represents signal intensity. n = 6. AUC, area under the curve. f, GSEA of selected GO processes on choline vs control treated human HSPCs. P-adjusted value. n = 3. g, Volcano plot of transcriptome depicting SREBF1 (left) and SREBF2 (right) target genes in choline vs control treated HSPCs by decoupleR. Color shows the regulation of targets by the respective TF. P-adjusted value. h, i-cisTarget prediction of TF regulatory motifs in up-regulated genes in choline vs control treated human HSPCs. TF logo is depicted. NES, Normalized Enrichment Score. i, Apoptosis quantification in human CD34+ cells after 48 h choline treatment using Annexin V and DAPI staining. j, GSEA of selected GO processes on choline vs control treated human aged HSPCs. P-adjusted value. n = 3. (b) Data are presented as mean ± SD. (c, e, f and i) n indicates the number of biological replicates per condition. For (c–e), 2 independent experiments were performed.
