NFAT5-deficient HSCs exhibit an altered transcriptome with enhanced MPP signatures. (A) MSigDB hallmarks enriched in the set of genes differentially upregulated (left panel) or downregulated (right panel) in HSCs sorted via FACS 13 weeks after bone marrow transplant (BMT) from Nfat5^fl/fl Vav-Cre or littermate WT mice (n = 3). See supplemental Figure 6A for the sorting strategy. (B) MSigDB hallmarks upregulated or downregulated in WT MPPs vs WT HSCs 13 weeks after BMT. MSigDB hallmarks analysis was done with Enrichr. (C) Venn diagrams of the number of differentially expressed genes upregulated (left) and downregulated (right) in NFAT5-deficient vs WT HSCs that were also upregulated or downregulated in WT MPPs vs HSCs after BMT. The bottom panels show the Enrichr analysis of MSigDB hallmarks enriched in the set of MPP-like genes upregulated (left panel) or downregulated (right panel) in NFAT5-deficient vs WT HSCs. (D) Enrichment in short-term HSC (ST-HSC) and early progenitor signatures (from MSigDB C2-CGP MM676 and MM1012 data sets respectively) in the set of genes upregulated after BMT in NFAT5-deficient (knockout [KO]) vs WT HSCs, and in WT or NFAT5-deficient (KO) MPP vs HSC. MSigDB were analyzed with gene set enrichment analysis. (E) Venn diagrams of the number of differentially expressed genes upregulated (left) and downregulated (right) in NFAT5-deficient (Nfat5^fl/fl Vav-Cre mice) vs WT HSCs that were also upregulated or downregulated, respectively, in WT MPPs vs HSCs in steady-state conditions. The fold-change (FC) values for the pairwise comparisons shown in the Venn diagrams in panels C,E were ≥2, with P values < .05. adj P, adjusted P value; CS, combined score; FDR, false discovery rate; NES, normalized enrichment score.