Fig. 1
Over-expression of wild-type IDH2 in AML and its effect on leukemia cell proliferation. a Comparison of IDH2 mRNA levels in AML (n = 80) and normal cells (monocytes and neutrophils, n = 6), using the leukemia dataset (Stegmaier Leukemia datasets) available in the Oncomine database. b IDH2 mRNA levels in primary AML samples (n = 204) in comparison with normal cells (hematopoietic stem cells, n = 6; metamyelocytes, n = 3; band cells, n = 3; polymorphonuclear cells, n = 3; monocytes, n = 4) available in the BloodSpot datasets. c Western blotting of IDH2 in normal human peripheral blood mononuclear cells (PBMC #1–#3), AML cell lines and PBMCs from AML patients with wt-IDH2 (AML#1–#8), β-actin was used as a loading control. d Relative mRNA level of IDH2 in U937 and ML-1 cells transfected with control shRNA (shRNA-Ctrl) or IDH2 shRNA (shIDH2#1, shIDH2#2) was measured by RT-qPCR. e Western blotting analysis of IDH2 protein in U937 and ML-1 cells transfected with shRNA-Ctrl or IDH2 shRNA. f, g Measurement of cell proliferation in U937 and ML-1 cells expressing shRNA-Ctrl or IDH2 shRNA. h Relative IDH2 mRNA level in HL-60 cells transfected with control vector (Vector) or IDH2 over-expression vector (IDH2OE). Expression of mRNA was measured by RT-qPCR. i Western blotting analysis of IDH2 protein levels in HL-60 cells transfected with control vector or IDH2 over-expression vector. j Comparison of cell proliferation in HL-60 cells transfected with control vector or IDH2 over-expression vector. k, l Colony formation assay in U937 and ML-1 cells expressing shRNA-Ctrl or IDH2 shRNA. Colonies were counted under inverted microscope. m Comparison of percentage of Annexin V/ PI negative cells in U937 and ML-1 transfected with shRNA-Ctrl or IDH2 shRNA for 48 h. The original flow cytometry analysis data are presented as Additional file 3: Fig. S1h. n = 3, mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001