Transcriptome analysis of hematopoietic progenitors reconstituted upon transplant of IFNAR1- and NFAT5-deficient HSCs. (A) Proportion and number of donor-derived LSK cells, HSCs and MPPs in bone marrow of mice that received transplantation with HSC donor cells (200 HSCs + 3 × 10⁵ CD45.1 Sca1^neg helper cells as in supplemental Figure 4A) of the indicated genotypes, and used for the RNA-sequencing analysis below. Results are shown as mean ± SEM. Statistical significance (unpaired t test) is shown for comparisons between the indicated genotypes. ∗P < .05, ∗∗P < .01. Transplants were done with 1 donor of HSCs of each genotype transplanted in 6 recipient mice. Equal numbers of FACS-sorted HSCs and MPPs from 6 recipient mice per genotype were pooled and processed for RNA sequencing. (B) Venn diagrams illustrating the numbers of differentially upregulated (FC ≥2.5) MPP-enriched (WT MPPs vs WT HSCs) genes in NFAT5-deficient (Nfat5 KO) vs WT HSCs; IFNAR1-deficient (Ifnar1 KO) vs WT HSCs; and double-deficient (DKO) vs WT HSCs. (C) Main MSigDB hallmarks enriched in the set of genes differentially upregulated (FC ≥2) in the indicated HSC comparisons. MSigDB hallmarks analysis was done with Enrichr. (D) Relative upregulation of IFN response genes in the MSigDB hallmarks database in pairwise comparisons of HSCs of the indicated genotypes. (E) Heat map showing the differential expression of a subset of 29 IFN-response genes in the comparisons indicated. This subset comprises genes annotated as IFN responsive in MSigDB and/or Interferome database, within the set of genes differentially upregulated (411 genes, FC ≥2) in NFAT5-deficient (Nfat5 KO) vs NFAT5 and IFNAR1 double-deficient HSCs (DKO). adj P, adjusted P value; CS, combined score.