NR4A1/3 suppress an inflammatory proliferative response in HSCs. (A) Flow cytometry quantification of IFNγ levels in HSCs. (B) Sca-1 frequency in Lin⁻ cells. (C-D) Flow cytometry quantification of protein levels in HSCs of (C) p-STAT1, p-AKT, and (D) p-ERK1/2 and MYC. (E) Flow cytometry quantification of phospho-p65 levels in HSCs. (F) Luciferase reporter assay after cotransfection of NR4A3 of GFP expression vectors with a luciferase reporter vector driven by a 5× NF-κB response elements in 32Dcl3 cells. (G-H) ChIP-Seq overlap between NR4A1 in Kasumi-1 cells and NF-κB in TNF-α-treated GM10847 cells in the (G) TNF locus and (H) IL6 locus. (I) ChIP-qPCR analysis of NR4A1 binding to NF-κB-regulated inflammatory cytokines in expanded human CD34⁺ bone marrow cells. (J) ChIP-Seq overlap between NR4A1 in Kasumi-1 cells and NF-κB in TNF-α-treated GM10847 in the IL1B +24 Kb enhancer. NR4A1 ChIP-Seq profile was extracted from Duren et al.³⁷  NF-κB ChIP-Seq profile was extracted from the publically available ENCODE dataset.⁶⁶  All flow cytometry analysis in A-E were performed at 4 days after first tamoxifen treatment. Results expressed as mean ± SD, n = 4 (A,G-I) or 3 (B-C). *P ≤ .05; **P ≤ .01; ***P ≤ .001.