Fig. 6: Sox4- induced cell state provides competitive advantage to Tet2 KO primitive cells.

a Schematics representing the experimental workflow for generating PVA-expanded HSPCs (Sox4 OE and Tet2 KO) and relative molecular analyses. b Flow cytometry analysis showing the distribution of the indicated primitive population after prolonged expansion of mouse HSC with PVA-containing medium for 3 weeks. n = 4, Data from 2 independent experiments are included. One-way Anova with Turkey’s multiple comparison test; Exact p-values are indicated on the figure. Data are presented as mean values +/- SEM. c Unsupervised clustering annotation of in vitro expanded mouse HSPC and analyzed by SHARE-seq (d) Selected pathways from GSEA analysis comparing gene expression data from HSC cluster from Fig. 6c. NES: Normalized enrichment score (e) Untargeted metabolomic analysis showing deregulated pathways comparing Sox4 OE Tet2 KO vs WT KSL HSC harvested from mice, n = 3. Statistical values obtained using Metaboanalyst 5.0 software¹¹⁸ (f) Schematics representing the experimental workflow for paired clonal tracking study on PVA-expanded HSPCs. g Clone size comparison of PVA-expanded HSC sister samples treated as indicated. Clone pairs Tet2 KO vs WT n = 97, Sox4 OE vs WT n = 130, Sox4 OE Tet2 KO vs WT n = 158. Blue line represents linear regression and highlights relative expansion of Sox4 OE Tet2 KO clones as compared to WT counterparts, (h) Percentage of CD45.2 cells measured at different time points in the PB after secondary BM transfer of cells from 6b. n = 5, Data from 1 independent experiment is included. Two-way Anova with Turkey’s multiple comparison test, Exact p-values are indicated on the figure. Data are presented as mean values +/- SEM. Source data are provided as a Source Data file.