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Figure 6. BMP4 induced MIXL1, an important survival axis and therapeutic target in AML(A) MIXL1 expression increased 2-fold in CD34+ HSPCs treated with BMP4. CD34+ HSPCs from three cord blood donors (unique donor number 47, 51, 60) were cultured with either 50 ng/ml BMP4 or 2 ng/ml TGF-β1 for 2 hours. MIXL1 transcript levels were quantified by RT-qPCR using 18S rRNA as normalization control. Error bars represent standard deviation between triplicates. *p < 0.05. (B) LDN-193189 at 3 μM was cytotoxic to OCI-AML2, ML3, KG1, and K562 cells but not U937 and HL60 cells. Each cell line was treated with vehicle or 3 μM LDN-193189 on day 0, and viability was measured every 24 hours by MTS assay in triplicate. (C) OCI-AML2, ML3, KG1, and K562 were sensitive to 200 nM LDN-193189, while the non–MIXL1-expressing lines U937 and HL60 were unaffected. Each cell line was treated in triplicate with 0–700 nM LDN-193189, with the drug or control medium replenished every 24 hours, for 4 days. Viability on day 4 was assayed by MTS assay. Absorbance was normalized to that of control samples treated with vehicle only. Image published in: Raymond A et al. (2014) Copyright: © 2014 Raymond et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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