HEK cells were plated in 12 or well plates. Protein lysates were obtained using urea lysis buffer 0. Lysates were sonicated and analyzed by 7. The anti-P-S antibody was raised on a service basis by Moravian Biotechnology and validated previously Luminescence was measured using a Synergy2 microplate reader BioTek. Sequences for one-to-one orthologs for each Class F receptor in human were downloaded from Ensembl 84 for all species with the exception of S. Statistical and graphical analysis were performed using Graph Pad Prism software. Concentration-response curves of BRET data were fit using three, four parameter or bell-shaped non-linear regression.
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