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CryoEM grids were prepared in the University of Virginia Molecular Electron Microscopy Core (MEMC) using a Vitrobot Mark IV with the blotting chamber at 4°C and 95 % relative humidity. Whatman #1 filter paper was used to blot grids using a blot force of 7 and blot time of 14 s. UltrAuFoil 1.2/1.3, 300 mesh grids (Quantifoil) were glow discharged in a Pelco EasiGlow for 60 s at 20 mA. A 2.1 µL aliquot of A2AAR-Gs was applied to the gold-film side of each grid and vitrified in liquid ethane cooled by liquid nitrogen.
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