GPCR Activation Monitoring Protocol

All materials were purchased from Sigma-Aldrich (Gillingham, UK) unless otherwise stated. FuGENE-HD, ONE-Glo™ luciferase and HaloTag® AF488 ligand were purchased from Promega Corporation (Wisconsin, USA). Opti-MEM was purchased from Thermo-Fisher Scientific (Massachusetts, USA). SNAP-Tag® AlexaFluor 647 (AF647) and AlexaFluor 488 (AF488) were purchased from New England BioLabs (Massachusetts, USA). VEGF₁₆₅a was purchased from R&D Systems (Abingdon, UK). VEGF₁₆₅a-TMR was produced as described by Kilpatrick et al. (2017) (link). BODIPY-CGP12177-TMR (Baker et al., 2003 (link)) was purchased from Molecular Probes (Oregon, USA). CGP12177 and ICI118551 were purchased from Tocris Bioscience (Bristol, UK). The cDNA construct expressing β-arrestin2-Venus-YFP was provided by Dr KDG Pfleger and generated as described in Kocan et al. (2008) (link).