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. 2015 Feb 12;106(3):227–236. doi: 10.1111/cas.12593

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© 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Functional domains of mixed lineage leukemia (MLL) required for interaction with PU.1 and for PU.1-mediated activation of macrophage colony-stimulating factor receptor (Csf1r) promoter. (a) PU.1 binding and PU.1-mediated Csf1r promoter activity of MLL deletion mutants. The PU.1-, menin-, and LEDGF-interacting domains and the results for interaction with PU.1 and PU.1-mediated transactivation of Csf1r-luc are indicated. ND, not determined. (b,c) Pu.1 binding. 293T cells were co-transfected with WT or mutants of HA-tagged MLL and FLAG-tagged PU.1 (b), or with WT or mutants of FLAG-tagged MLL and HA-tagged PU.1 (c). Anti-FLAG antibody immunoprecipitates (IP:FLAG) or cell lysates (Input) were subjected to immunoblotting with anti-HA or anti-FLAG antibodies. (d) PU.1-mediated Csf1r promoter-driven transcription. SaOS2 cells were transfected with the Csf1r–luciferase construct and PU.1, together with deletion mutants of MLL. Luciferase activity was analyzed 24 h after transfection. Error bars represent SD (n = 3).