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To quantify SNAP/FLAG-hGLP-1R association with clathrin, circular ROIs surrounding individual well defined puncta of fluorescence (5-50 per cell) were selected on CLC-GFP images and transferred to the corresponding SNAP ones using a Fiji macro and average fluorescence per pixel was measured in a central circle (c) of diameter 5 pixels (0.55 m) at each puncta location, in a surrounding annulus (a) with an outer diameter of 7 pixels (0.77 m), and in an area not containing any cell as background (bg)

Sheka AC, Adeyi O, Thompson J, Hameed B, Crawford PA, Ikramuddin S