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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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Cell adhesion on fibronectin triggers GRB2-MPZL1 complex formation at the plasma membrane. (a) HEK293T cells expressing FLAG-GFP or GRB2 were seeded on plastic or fibronectin for ten minutes before lysis. PV treated cells were used as a positive control. FLAG APs were performed for each condition and analyzed by Western blot as indicated. (b) HCC1954 cells were seeded on polylysine or fibronectin for 15 minutes before fixation. Immunofluorescence against GRB2 and MPZL1 was performed to assess the cellular localization. Confocal images of the z-axes (indicated by yellow lines) are shown on the left side (y-z axis) or on top (x-z axis) of the panels showing merged images, for which nuclei were stained with DAPI. Representative images are presented (scale bar = 5 μm). (c) HCC1954 cells were depleted of endogenous PTPN11 using DsiRNAs. (d) PTPN11-depleted cells were seeded on fibronectin for 15 minutes prior to fixation and immunofluorescence. Confocal images of the z-axes (indicated by yellow lines) are shown on the left side (y-z axis) or on top (x-z axis) of the panels showing merged images, for which nuclei were stained with DAPI. Representative images are shown (scale bar = 5 μm).