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. 2018 Dec 10;7:e42908. doi: 10.7554/eLife.42908

Figure 2. Effect of artificially enforced GPCR-GIRK co-localization.

Figure 2.

(A) A schematic representation of GPCR-GIRK concatemer constructs. GIRK was directly fused to the C-terminus of GPCRs. A cleavable signal peptide and a Halo tag were added to the N-terminus of each concatemer. Additionally, a SNAP tag was added to the C-terminus of each concatemer. (B) Western-Blot analysis of GPCR-GIRK concatemer constructs. HEK-293T cells were transiently transfected with either M2R-GIRK or β2AR-GIRK concatemers. The expected size of these concatemers is ~150 kDa. (C) (D) Representative voltage-clamp recordings of HEK-293T cells transiently transfected with M2R-GIRK concatemers or β2AR-GIRK concatemers. Membrane potential was held at −80 mV. 10 µM ACh or Iso was applied as indicated. (E) Validation of the function of β2AR-GIRK concatemers. HEK-293T cells expressing β2AR-GIRK concatemers were treated with 10 µM propranolol (Pro) or isoproterenol (Iso), and intracellular cAMP levels were quantified (N = 3, ±SD).