![IL-1α–exposed Tet2+/−HSCs upregulate proliferation and maintain self-renewal transcriptomic signatures. (A) Experimental design. (B) Principal component analysis (PCA) plot of WT HSCs treated with PBS (n = 5) or IL-1 (n = 4) and Tet2+/− HSCs treated with PBS (n = 3) or IL-1α (n = 3) based on regularized log gene-level counts. (C) Venn diagram depicting differentially expressed genes (DEGs; −1 > log2[fold change {FC}] > 1; false discovery rate [FDR] < 0.05) in HSCs exposed to IL-1α or PBS, which are unique to WT, unique to Tet2+/−, or shared between the 2 groups. (D) Overrepresented GO BP terms (maximum of 10 terms) for upregulated DEGs, shared between WT and Tet2+/− HSCs exposed to IL-1α. GO BPs are displayed in ascending order, according to –logFDR value. (E) Upregulated shared DEGs (maximum of 20 genes) present in indicated overrepresented GO BPs. (F) Co-occurring transcription factors (TFs; maximum of 10) with shared UP DEGs. TFs are displayed in ascending order, according to –logFDR value. Boxplot Spi1 gene expression values (fragments per kilobase million [FPKMs]) in indicated groups (right). (G) Overrepresented GO BP terms (maximum of 10 terms) for upregulated DEGs unique to Tet2+/− HSCs exposed to IL-1α. GO BPs are displayed in ascending order, according to –logFDR value. (H) Upregulated DEGs (maximum of 20 genes) unique to Tet2+/− IL-1α group present in indicated overrepresented GO BPs. (I) Co-occurring TFs (maximum of 10) with upregulated DEGs unique to Tet2+/− IL-1α group. TFs are displayed in ascending order, according to –logFDR value. Boxplot of E2f1 gene expression values (FPKMs) in indicated groups (right). (J) Heat map showing expression of genes implicated in HSC self-renewal. Column Z-scores are normalized. All genes displayed are significantly downregulated between PBS and IL-1α conditions, independently of HSC genotype (first gene set); PBS and IL-1α condition in WT HSCs (second gene set); and PBS and IL-1α condition in Tet2+/− HSCs (third gene set). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 by a 1-way analysis of variance with Tukey correction. RNA-seq, RNA sequencing.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/141/8/10.1182_blood.2022016835/8/blood_bld-2022-016835-gr5b.jpeg?Expires=1721550049&Signature=jeOhqrKi5ExVpcAL5YE8v4K572PNIpBKmXm6vg~n~~o~CKtXyvGQ5Nh61o2Sue3edHrBsB7lNzLBQxbP3czHXM~71TU46M~ZMR9p60vBRgextxo2rURLeK2NCom~-Tk0OlU1aa8SzKWuXLYyG4SMLYeKWdDgKe94t-wggTf9wTbz5ybmKaADwSqOZp9HuosZklHrpN6Oudv6N2Q8Ey8yAvIj5P7dVftIAdS~4KVR6PhgwpiHqh9VAxzdc3aw~uYAr7lf934X0VW29dB0h88tFjKxG-jmrt8q8P37xssOAvyOD6ywff1GcgwUpIKw9KdlLJHc21lsVhpqlwdVVbLnSg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
IL-1α–exposed Tet2+/−HSCs upregulate proliferation and maintain self-renewal transcriptomic signatures. (A) Experimental design. (B) Principal component analysis (PCA) plot of WT HSCs treated with PBS (n = 5) or IL-1 (n = 4) and Tet2+/− HSCs treated with PBS (n = 3) or IL-1α (n = 3) based on regularized log gene-level counts. (C) Venn diagram depicting differentially expressed genes (DEGs; −1 > log2[fold change {FC}] > 1; false discovery rate [FDR] < 0.05) in HSCs exposed to IL-1α or PBS, which are unique to WT, unique to Tet2+/−, or shared between the 2 groups. (D) Overrepresented GO BP terms (maximum of 10 terms) for upregulated DEGs, shared between WT and Tet2+/− HSCs exposed to IL-1α. GO BPs are displayed in ascending order, according to –logFDR value. (E) Upregulated shared DEGs (maximum of 20 genes) present in indicated overrepresented GO BPs. (F) Co-occurring transcription factors (TFs; maximum of 10) with shared UP DEGs. TFs are displayed in ascending order, according to –logFDR value. Boxplot Spi1 gene expression values (fragments per kilobase million [FPKMs]) in indicated groups (right). (G) Overrepresented GO BP terms (maximum of 10 terms) for upregulated DEGs unique to Tet2+/− HSCs exposed to IL-1α. GO BPs are displayed in ascending order, according to –logFDR value. (H) Upregulated DEGs (maximum of 20 genes) unique to Tet2+/− IL-1α group present in indicated overrepresented GO BPs. (I) Co-occurring TFs (maximum of 10) with upregulated DEGs unique to Tet2+/− IL-1α group. TFs are displayed in ascending order, according to –logFDR value. Boxplot of E2f1 gene expression values (FPKMs) in indicated groups (right). (J) Heat map showing expression of genes implicated in HSC self-renewal. Column Z-scores are normalized. All genes displayed are significantly downregulated between PBS and IL-1α conditions, independently of HSC genotype (first gene set); PBS and IL-1α condition in WT HSCs (second gene set); and PBS and IL-1α condition in Tet2+/− HSCs (third gene set). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001 by a 1-way analysis of variance with Tukey correction. RNA-seq, RNA sequencing.
