Fig. 2.

JMJD3 bears oncorepressor activity in mouse AML models. a, b Western blotting (a) or qRT-PCR (b) assay on Jmjd3 protein or mRNA level of GFP⁺ murine AML BM myeloid cell samples expressing PML-RARα, AML1-ETO9a, or MLL-AF9, and a normal sorted CD11b⁺ BM myeloid cell sample. c AML1-ETO, PML-RARα, or MLL-AF9 was transduced into normal c-Kit⁺ BM cells by retroviral infection, and the mRNA level of JMJD3 of the transduced GFP⁺ cells was measured by qRT-PCR. d Experimental protocol for testing the in vivo proliferative capacity of GFP⁺ BM leukemic cells transduced with YFP⁺ vector control or JMJD3-expressing vector in FVB/NJ or C57BL/6J mice. e GFP⁺ murine AML BM cells expressing PML-RARα, AML1-ETO9a, or MLL-AF9 were transduced with YFP⁺ empty vector or JMJD3-expressing vector, and the mRNA levels of mouse Jmjd3 or human JMJD3 were measured by qRT-PCR. f The percentages of PML-RARα (left panel) or AML1-ETO9a (right panel) GFP⁺YFP⁺ cells in the peripheral blood of syngeneic recipients after they had been transduced with YFP⁺ empty vector or JMJD3-expressing vector. g Kaplan–Meier curves are shown for syngeneic mice transplanted with GFP⁺ AML (left panel, PML-RARα⁺ cells; right panel, AML1-ETO9a⁺ cells) BM cell transduced with YFP⁺ empty vector or JMJD3-expressing vector. h The percentages of MLL-AF9⁺ GFP⁺YFP⁺ cells in the peripheral blood (left panel) and the survival (right panel) of the syngeneic recipients transduced with YFP⁺ empty vector or JMJD3-expressing vector. i, j Flow cytometric analyses of Gr-1 expression (i) or CD11b expression (j) on PML-RARα⁺ (left panel) or AML1-ETO9a⁺ (right panel) leukemia BM cells transduced with YFP⁺ empty vector or JMJD3-expressing vector and cultivated for 72 h. k Flow cytometric analyses of Annexin V staining on PML-RARα⁺ (left panel) or AML1-ETO9a⁺ (right panel) leukemia BM cells transduced with YFP⁺ empty vector or JMJD3-expressing vector. l–n Flow cytometric analyses of Gr-1 expression (l), CD11b expression (m), or Annexin V staining (n) on MLL-AF9⁺ BM cells transduced with YFP⁺ empty vector or JMJD3-expressing vector in vivo. Data are shown as the mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001