AML inhibits osteogenesis through interference in WNT/β-catenin pathway. (A) GSEA enrichment plot of GO_WNT_ACTIVATED_RECEPTOR_ACTIVITY of bone marrow MSPCs from AML patients and controls (NES, −1.55; P = .021; false discovery rate q = 0.114). (B) Representative overlay histogram of cytoplasmic β-catenin fluorescence intensity in HuMSPCs after mono- or coculture with healthy PBMNCs (controls) or indicated AML cell lines. (C) Mean fluorescence intensity (MFI) of cytoplasmic β-catenin levels in HuMSPCs after mono- or coculture with the indicated cells, normalized to HuMSPCs (representative of 5 independent experiments). (D) Colorimetric detection of alizarin red staining using absorbance at 450 nm to quantify calcium deposition in SaOS2 cells transduced with either empty vector or different CTNNB1 short hairpin RNAs after 7 days of induction of mineralization (representative of 4 independent experiments). (E) Colorimetric detection of alizarin red staining using absorbance at 450 nm to quantify calcium deposition in HuMSPCs after 14 days of induction of mineralization in the presence of healthy PBMNCs or different AML cell lines with either 10 µM SKL2001 or dimethyl sulfoxide (DMSO) (representative of 8 independent experiments; result is normalized to HuMSPCs with DMSO). (F) MFI of cytoplasmic β-catenin levels in HuMSPCs after mono- or coculture in the presence of healthy PBMNCs or different AML cell lines with either 10 µM SKL2001 or DMSO (representative of 5 independent experiments; result is normalized to HuMSPCs with DMSO). (G) Colorimetric detection of alizarin red staining using absorbance at 450 nm to quantify calcium deposition in SaOS2 cells transduced with either empty vector or CTNNB1 overexpressing plasmids after 7 days of induction of mineralization in the presence of different AML cell lines (representative of 3 independent experiments). Data are shown as mean ± SEM. *P < .05; **P < .01; ***P < .001 (determined by unpaired Student t test).