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. 2017 Sep 14;7:11514. doi: 10.1038/s41598-017-11876-9

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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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The tyrosine phosphatase PTPN11 is required as a scaffold for the GRB2/MPZL1 interaction. (a) HEK293T were treated with PV and lysates were used to perform APs using the indicated antibodies, followed by a Western blot analysis. (b) HEK293T cells were depleted of endogenous PTPN11 using DsiRNAs and transiently transfected with FLAG-GFP or GRB2 transgenes. FLAG APs were performed on cell lysates following PV treatment and analyzed by Western blot. (c) Cells obtained as in (b) were transfected with a GFP-tagged murine PTPN11 that is DsiRNA-resistant. Cells were treated with PV and FLAG APs were performed for each condition, prior to Western blotting analysis. (d) HEK293T cells were transfected with FLAG-GFP, MPZL1 (WT or non- phosphorylatable Y/F point mutants), treated with PV and utilized for FLAG APs - Western blotting.