Fig. 4.

JMJD3 induces the expression of key myelopoietic regulators in AML cells. a Heatmap showing the differentially expressed genes between HL-60 cells transduced with control or JMJD3-expressing vector (fold change ≥2, p < 0.05). b–g GSEA of the expressing profile of HL-60 cells transduced with control or JMJD3-expressing vector using a senescence-associated upregulated signature (b), a p53 pathway-associated signature (c), an innate immune system-associated signature (d), a NF-κB pathway-associated signature (e), a leukemic stem cell (LSC)-associated upregulated or downregulated signature (f), and a myeloid cell development-associated upregulated or downregulated signature (g). h Differential expression analyses of HL-60 cells upon overexpression of JMJD3 (fold change ≥2, p < 0.05) (top panel), and validation of the mRNA levels of a number of genes by qRT-PCR (bottom panel). FPKM, fragments per kilobase of transcript per million fragments mapped. i Differential expression analysis upon knockout of JMJD3 in HL-60 (fold change ≥1.5, p < 0.05) (top panel), and validation of the mRNA levels of a number of genes by qRT-PCR (bottom panel). j Two primary AML blasts and the NB4 cells were transduced with empty vector or JMJD3-expressing vector, and the mRNA levels of C/EBPβ or RIPK3 were measured by qRT-PCR. k NOD/SCID mice injected with HL-60 cells transduced with vector control, JMJD3-, or H1390A mutant-expressing vector, and qRT-PCR assay on the C/EBPβ or RIPK3 mRNA level of GFP⁺ HL-60 cells isolated from the BM at 35 days after transplantation. l GFP⁺ murine AML BM cell samples from PML-RARα-, AML1-ETO-, or MLL-AF9-expressing transgenic mice were transduced with empty vector or JMJD3-expressing vector, and the mRNA levels of C/EBPβ or RIPK3 were measured by qRT-PCR. m Correlated mRNA levels of C/EBPβ or RIPK3 with those of JMJD3 in 179 AML patients (raw data from TCGA database). n–p HL-60 cells transduced with vector control or JMJD3-expressing vector were further treated with NC siRNA, C/EBPβ siRNA, or RIPK3 siRNA. Flow cytometric analyses of CD11b expression (n), annexin V staining (o), and cell cycle status (p). Data are shown as the mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001