SGEF’s N terminus interacts with Dlg1 GUK domain. (A) Schematic representation of SGEF and Dlg1 constructs used in this study. (B–F) Lysates from HEK293FT cells expressing the indicated constructs were immunoprecipitated using GFP antibodies (GFP-trap nanobodies). In all experiments, the precipitates were immunoblotted with anti-GFP antibodies to detect the immunoprecipitated protein and with anti-myc to detect the interacting partner. (G) Sequence alignment comprising the Dlg1 binding domain of SGEF in vertebrates (aa 301–350 for human).