Extended Data Fig. 10: Validation of multipotency-associated genes.

a, Representative gating schemes for FACS-purification of mouse hematopoietic subsets analyzed in Fig. 2i (HSCs/MPPs, CMPs, and CLPs from bone marrow and T/B cells from peripheral blood). KLS, Kit⁺ Lin⁻ Sca1⁺ multipotent cell subset consisting of HSCs and MPPs (multipotent progenitors); KL, Kit⁺ Lin⁻ Sca1⁻ subset devoid of multipotent cells; CMP, common myeloid progenitor; CLP, common lymphoid progenitor. b, Bar plots showing biological replicates and controls for quantitative PCR experiments, related to Fig. 2i. Each gene is shown normalized to the maximum mean expression across all groups. Top: markers of HSC/MPP (Hoxb5, Fgd5, Procr), progenitors (Cd34), and differentiated lineages (Cd8a, Cd19). Bottom: unsaturated fatty acid (UFA) synthesis genes identified as markers of multipotency by CytoTRACE 2 (Fads1, Fads2, and Scd2). Actb was used as an internal control. Error bars reflect s.e.m. (standard error of the mean) across three biological replicates. c-d, Same as Fig. 2j but shown for mouse duodenum (c) and ileum (d). Scale bars, 50 µm (top) and 10 µm (bottom). e, Quantification of mRNA hybridization signal in multipotent and unipotent/differentiated zones of mouse jejunum (left, corresponding to images in Fig. 2j), duodenum (center, corresponding to confocal images in c), and ileum (right, corresponding to images in d). Multipotent zones are divided as previously described⁴⁰ on the basis of cell location from the crypt base, with red and green regions expected to be most enriched in Lgr5 and Fgfbp1, respectively. Bars represent the mean fluorescence intensity per zone, with error bars denoting s.e.m. (n = 20 paired crypts and villi from each intestinal region [jejunum, duodenum, ileum] pooled from two mice, with a total of 10 paired crypts and villi per region per mouse). Statistical significance was determined by a two-sided paired t test, with the resulting p-values adjusted by the Benjamini-Hochberg method separately applied to jejunum, duodenum, and ileum samples (*Q < 0.05; **Q < 0.01, ***Q < 0.001, ****Q < 0.0001).