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. 2016 Apr 4;113(16):4356–4361. doi: 10.1073/pnas.1516767113

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Fig. 1. — Gip1 is an interactor of trimeric G protein. (A) TAP using Flag×2-GFP-Gβ (F₂G-Gβ) in vegetative growth (v) and before (0'') and 30 s after (30'') cAMP stimulation. Silver staining showed F₂G-Gβ copurified with Gip1 along with ElmoE, Gα, and Gγ. (B) Gip1-GFP-Flag (Gip1-GFPF) and Gα4-GFP-Flag (Gα4-GFPF) proteins were expressed in WT cells and immunoprecipitated (IP) with α-Flag antibody from WCE. Copurification of Gβ (Upper) and GFP-Flag (Lower) was evaluated by immunoblotting. Gα4-GFPF was used as a positive control and confirmed the coprecipitation of Gβ. (C) Full-length Gip1 and its C and N termini tagged with GFP-Flag (Gip1-GFPF) were immunoprecipitated with α-Flag antibody from WCE. Copurification of Gβ (Upper) and GFP-Flag (Lower) was evaluated by immunoblotting. (D) Cells were starved on nonnutrient DB agar. Gip1^OE cells are WT cells with full-length Gip1 overexpression.