Fig. 5
Regnase-1-deleted HSPCs show a phenotype and gene expression profile similar to leukemia. a Representative field of Wright-Giemsa-stained BM smears of 8-week-old control (fl/fl) and Regnase-1-KO (Δ/Δ) mice. The scale bar shows 30 μm. b Quantification of frequencies of blasts and other MNCs in PB on Wright-Giemsa-staining (n = 3 per group; 3 independent experiments). c Quantification of abnormal cells in the PB. d Peripheral white blood cell counts in blood samples from 8-week-old control (fl/fl) or Regnase-1-KO (Δ/Δ) mice (n = 3 per group). e Wright-Giemsa staining of CD34− HSCs. The scale bars represent 30 μm. f Scatter-plot representation of the transcriptional differences between control Reg1flox/flox (fl/fl) and Vav1-iCre; Regnase-1flox/flox(Δ/Δ) CD34− HSCs. The color indicates >2-fold differential gene expression (n = 3 per group and n = 2 per group; 2 independent experiments; the average of the two is shown). g Gene set enrichment analysis comparing CD34− HSCs from control (fl/fl) and Regnase-1-KO (Δ/Δ) mice with the association between genes upregulated following Regnase-1 deletion and Pu.1 deletion. h Human HSC signatures of AML compared with Regnase-1-deficient CD34− HSCs. The normalized enrichment scores (NES), P-value and q-value (FDR) are indicated on each plot. i Kaplan-Meier plot of AML patient survival data based on similarity of gene expression profiles of loss of Regnase-1. P values were calculated by the logrank test. j Regnase-1 mRNA expression (upper panel) and proliferation (lower panel) of THP1 and HL60 leukemic cells transfected with mouse Regnase-1 cDNA expression vector (n = 3 per group; 3 independent experiments). Error bars indicate mean ± SD. *p < 0.05; **p < 0.01, ***p < 0.005, log rank test in i, two-sided t-test in c, d, j
