Fig. 4: Lipid molecule in ADGRF1.

a, Lipid-binding site in ADGRF1. The ADGRF1–miniG_s structure is shown in cartoon representation. The receptor is also shown as surface. The lipid LPC is shown as yellow sticks. The receptor residues involved in lipid binding are shown as blue sticks. b, High-resolution tandem mass spectrometry (MS/MS) spectra of two LPC molecules specifically associated with ADGRF1. Their experimental spectra matched with the reference spectra recorded in the Lipid-Blast database. c, Basal activity of wild-type (WT) and mutant versions of ADGRF1, measured by cAMP accumulation assay. Data are presented as a percentage of wild-type activity and are shown as mean ± s.e.m. (bars) from at least five independent experiments performed in technical triplicate with individual data points shown (dots). *P < 0.05, **P < 0.001, ***P < 0.0001 by one-way analysis of variance followed by Dunnett’s post-test compared to the response of wild type. ^#Low surface expression level (less than 40% of wild-type expression level). Extended Data Table 2 provides detailed independent experiment numbers (n), P values and expression level. d, A8-induced G_s and G_i activation of ADGRF1. Data are shown as mean ± s.e.m. from at least four independent experiments performed in technical duplicate. Extended Data Table 3 provides detailed independent experiment numbers (n), P values, statistical evaluation and expression level.