Fig. 6: Sucnr1 preserves transcriptional programs characteristic of HSPC.

A, B RNA sequencing in fluorescence-activated cell sorting (FACS)-sorted Lin⁻Sca-1⁺c-Kit⁺ (LSK) from bone marrow (BM) of 27–30 week-old C57BL6/J wild-type (WT, n = 4) and Sucnr1 knock-out (Sucnr1-KO, n = 3) mice. A Volcano plot. Red and blue dots represent differentially expressed genes with adjusted p˂0.05 and absolute log₂FC > 1. Fold-change, FC. B Gene Set Enrichment Analysis (GSEA). HSC, hematopoietic stem cells; ROS, reactive oxygen species. C Digital droplet PCR (ddPCR) mRNA expression of S100a8 (n = 10 per group), S100a9 (n = 10 per group), Il1b (n = 10 WT, n = 9 Sucnr1-KO), Procr (n = 8 WT, n = 6 Sucnr1-KO) and Hoxa9 (n = 8 WT, n = 6 Sucnr1-KO) relative to Gapdh in FACS-sorted LSK of 35–44 week-old mice. D ddPCR mRNA expression of Procr (n = 10 LT, ST and n = 8 MPP WT, n = 9 LT, ST and n = 8 MPP Sucnr1-KO) and Hoxa9 (n = 10 LT, ST and n = 8 MPP WT, n = 9 LT, ST and n = 7 MPP Sucnr1-KO) relative to Gapdh in FACS-sorted LSK subsets of 30–45 week-old mice; long-term HSC (LT, LSK CD34⁻Flt3⁻), short-term HSC (ST, LSK CD34⁺Flt3⁻) and multipotent progenitors (MPP, LSK CD34⁺Flt3⁺). E (Left) Intracellular pro-IL-1β in LSK-gated cells from total BM of 43 week-old WT and Sucnr1-KO mice (n = 3 per group) analyzed by FACS. Mean fluorescence intensity, MFI. (Right) Representative histograms in LSK-gated cells using spectral flow cytometry. F Pro-inflammatory IL-1β, IL-1α, MIP-1β, MCP-1, RANTES, GM-CSF levels in BM extracellular fluid (BMEF) from 40-45 week-old WT (n = 8) and Sucnr1-KO (n = 11) mice and IL-6 level in BMEF from 19 to 35 week-old WT (n = 12) and Sucnr1-KO (n = 13) male and female mice, analyzed by Bioplex. G Anti-inflammatory IL-9 and IL-13 levels in BMEF from 40-45 week-old WT (n = 8) and Sucnr1-KO (n = 9 IL-9, n = 8 IL-13) mice, analyzed by Bioplex. H Levels of S100A8 and S100A9 in BMEF of 41-46 week-old WT (n = 9) and Sucnr1-KO (n = 7) mice, analyzed by ELISA. Statistical analyses were performed with two-tailed Wald test with Benjamini-Hochberg correction for multiple comparisons (A), two-tailed Student’s t test (C–E, F except IL-1β, H), or two-tailed Mann–Whitney U test (F IL-1β, G). Significant p-values are reported. Data are biologically independent animals, and means ± SEM for bar plots. Source data and non-significant p-values are provided as a Source Data file.