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. 2017 Nov 30;7:16604. doi: 10.1038/s41598-017-16799-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Unidirectional regulatory loop of RUNX1-p53-CBFB. (a) RUNX1-depletion-mediated up-regulation of CBFB is attenuated by the additional knockdown of p53 (left panel). MV4-11 cells were lentivirally-transduced with the indicated combinations of shRNAs, and treated with 3 μM of doxycycline. Forty-eight hours after treatment, cell lysates were analyzed by immunoblotting with the indicated antibodies. GAPDH was used as a loading control (right panel). (b) p53-depletion-mediated decrease in the amount of RUNX1 is restored by ectopic expression of CBFB (left panel). MV4-11 cells were transduced with the indicated combinations of lentivirus vectors and treated with 3 μM of doxycycline. Forty-eight hours after treatment, cell lysates were analyzed by immunoblotting with the indicated antibodies. GAPDH was used as a loading control (right panel). (c) CBFB-depletion-mediated increase in the amount of p53 is attenuated by forced expression of RUNX1 (left panel). MV4-11 cells were transduced with the indicated combinations of lentivirus vectors and treated with 3 μM of doxycycline. Forty-eight hours after treatment, cell lysates were analyzed by immunoblotting with the indicated antibodies. GAPDH was used as a loading control (right panel). (d) The presence of unidirectional regulatory loop of RUNX1-p53-CBFB (upper panel). MV4-11 cells were exposed to 2 μM of Ro5-3335. At the indicated time points after treatment, cell lysates were analyzed by immunoblotting with the indicated antibodies. GAPDH was used as a loading control (lower panel). Signal Intensities of the indicated bands were quantitated by Image Lab software.