Fig 3. Validation of novel GCGR interactors by Co-IP/WB in CHO cells.
HA-tagged interactors and Flag-tagged GCGR were co-transfected into CHO cells, while cells only transfected with HA-tagged interactors were used as a control. Anti-Flag co-immunoprecipitation (Co-IP) and anti-HA Western blot were performed. A) Representative gels from the Co-IP/WB. I = Input proteins before Co-IP, W3 = 3rd wash, E = co-IP elutes. N = 3 per group. B) The relative binding strength for each interactor. For each of the interacting proteins, intensity of the elute/input band was quantified and expressed as a ratio to the lysate band (E:I ratio). The E:I ratio was used to estimate the percentage of input interactors binding to GCGR.