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. 2019 Nov 25;10:5351. doi: 10.1038/s41467-019-13259-2

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© The Author(s) 2019

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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Fig. 4 — EBP1 strongly interacts with HOXB-AS3 in NPM1mut AML cells. a–c Yields of a HOXB-AS3 RNA, b U1 RNA, and c GAPDH RNA in the eluates and the flow-through of lysates hybridized with U1 and HOXB-AS3-targeting probes. The RNA yield is depicted as a percentage of the amount of the respective transcript in the input sample. d Validation of HOXB-AS3-protein interactions via RNA-Immunoprecipitation (RIP) experiments with nine candidate proteins. Enrichment of each immunoprecipitate is compared with the respective IgG control (mouse or rabbit). *P < 0.05, **P < 0.005, ***P < 0.001, N.S., not significant. e Immunoprecipitation of the EBP1 protein in nuclear lysates of OCI-AML3 cells followed by western blotting (WB) for the EBP1 and NPM1 proteins. f Immunoprecipitation of the NPM1 protein in nuclear lysates of OCI-AML3 cells followed by WB for the EBP1 and NPM1 proteins. g–j Effect of HOXB-AS3 depletion (HOXB-AS3 KD) on the formation of the EBP1-NPM1 complex in OCI-AML3 cells: g EBP1 immunoprecipitation in scramble versus HOXB-AS3 KD-treated cells followed by WB for the EBP1 and NPM1 proteins. h Quantification of three independent experiments. i NPM1 immunoprecipitation in scramble versus HOXB-AS3 KD-treated cells followed by WB for the EBP1 and NPM1 proteins. j Quantification of three independent experiments. P values were calculated using paired two-sided t-tests. In the figures, heights of boxplots indicate mean values with standard deviation. Error bars indicate highest and lowest values in each population. Source data are provided as a Source Data file.