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. 2017 Oct 23;7:13741. doi: 10.1038/s41598-017-14096-3

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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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Removal of positive charge from residue⁶⁵ in hFFA2 affects response to agonists in G_q/11- and G_12/13-coupled TGFα shedding assays in a manner akin to inositol monophosphate assays. TGFα shedding assays were performed in response to varying concentrations of C3 (a,c,e,g) or compound 1 (b,d,f,h) in parental HEK293 cells (a,b), or those genome-edited to lack both Gα_q and Gα₁₁ (c,d), both Gα₁₂ and Gα₁₃, (e,f) or each of Gα_q, Gα₁₁, Gα₁₂ and Gα₁₃ (g,h) and transfected to express either wild type hFFA2 (dotted lines) or Lys⁶⁵Arg hFFA2 (○), Lys⁶⁵Ala hFFA2 (□) or Lys⁶⁵Glu (△) hFFA2.