IL27Ra+ HSCs are inflamed and TNF-α impairs HSC function via IL27Ra pathway. (A-B) These figures show the GSEA of inflammation-related genes in (A) Y+ (Young IL27Ra+ HSC) vs Y− (Young IL27Ra− HSC) and (B) O+ (Old IL27Ra+ HSC) vs O− (Old IL27Ra− HSC). FDR adjusted q values are provided. |NES| >0.3 and q < 0.05 represent significant difference. (C-D) The histogram shows read counts of 6 genes in (C) Y− (Young IL27Ra− HSC) and Y+ (Young IL27Ra+ HSC) and (D) O− (Old IL27Ra− HSC) and O+ (Old IL27Ra+ HSC). Data are shown as mean ± SEM. (E) The figures show the GSEA of inflammation-related genes in O+ (Old IL27Ra+ HSC) vs Y+ (Young IL27Ra+ HSC). FDR adjusted q values are provided. |NES| >0.3 and q < 0.05 represent significant difference. (F-H) Freshly isolated 100 HSCs from WT or IL27Ra−/− mice were treated by TNF-α (50 ng/mL) or control for 24 hours and then were transplanted into lethally irradiated recipients together with 2 × 105 competitor cells. Chimera in peripheral blood was evaluated every month until the fourth month. (F) Experimental design to evaluate HSC function in response to TNF-α treatment, (G) The line plots depict the peripheral blood chimera of recipients at the fourth month. N = 5-6 mice per group; data are shown as mean ± SEM. (H) This histogram depicts the lineage distribution of myeloid, B, and T cell 4 months after transplantation. Data are shown as mean ± SEM. (I) The schematic diagram showing the experimental design to evaluate the in vitro influence of TNF-α on myeloid differentiation in the context of IL27Ra status. The scatterplots depict the percentage of Gr1+Mac1+ cells upon TNF-α stimulation. Data are shown as mean ± SEM.