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. 2020 Oct 8;10:16809. doi: 10.1038/s41598-020-74032-w

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Characterization of PBX1 mutants. Panel A: Mapping of the heptad repeats of PBX1within PBC-A and PBC-B. Predicted alpha-helical regions are represented as yellow areas above the linear sequence. Boundaries of PBC-A, PBC-B and homeodomain are defined below the linearized sequence. Panel B: Time-resolved fluorescence immunoassay (TR-FIA)-measured binding of wild-type and mutant PBX1 to wild-type PREP1. The Y axis reports the binding to PREP1 of the PBX1 deletion mutants, normalized for the binding of wild-type PBX1 taken as 1. The results represent the average of three independent assays, + /− the standard deviation. The extent of deletions is indicated below the ordinates bar. Panel C: TR-FIA-measured binding of wild-type and mutant PBX1 to wild-type MEIS1. The Y axis reports the binding to MEIS1 of the PBX1 deletion mutants, normalized for the binding of wild-type PBX1 taken as 1. The results represent the average of three independent assays, + /− standard deviation. Panel D: TR-FIA-measured binding of wild-type and point mutants of PBX1 to wild-type PREP1. Single and multiple mutations analysed are indicated below the bars. PBC-A and PBC-B are mapped below the ordinates bar. The Y axis reports the binding to PREP1 of the PBX1 mutants, normalized for the binding of wild-type PBX1 taken as 1. The results represent the average of four independent assays, + /− the standard deviation. T-TEST was performed as described in the Methods section.