performed in singlicate, dose response curves performed in duplicate Error bars are mean +/ SEM GPCR Gastrointestinal Gastrointestinal N-acyl amide Genus Duodenum GPR119 N-acyl serinols lleum S1PR4 N-acy seria Ileum PTGIR lleum Colon PTGER4 Klebsiella Extended Data Figure 4. Combined analysis of protein and transcript expression of GPCR in the Table links GPCR, N-acyl amide, bacterial genus and the site where these co-occur in the gastrointestinal tract(colored). Based on protein expression data(Human Protein Atlas) GPRI19 is most highly expressed in the pancreas and duodenum, SIPR4 in the spleen and lymph node, G2A in the lymph node and appendix, PTGir in the lung and appendix and PTGER4 in the bone marrow and small intestine From gene expression data in the colon (GTEx dataset, N=88 patient samples from small intestine, 345 patient samples from colon) GPR132, PTGER4, and PTGir are all expressed alongside the N-acyl synthase genes known to encode metabolites that target these gPCr (Figure 1). In the gastrointestinal tract GPRI19 and SIPR4 are most highly expressed in the small intestine where 16S studies have identified bacteria from the genera Gemella and Neisseria. All known reference genomes (NCBI) from these genera contain N-acyl synthase genes that are highly similar(blastN,e value 2e-132)to those we found to encode GPR119 or SIPR4 ligands37,43,44 Nature. Author manuscript; available in PMC 2018 February 28performed in singlicate, dose response curves performed in duplicate. Error bars are mean +/ − SEM. Extended Data Figure 4. Combined analysis of protein and transcript expression of GPCR in the gastrointestinal tract Table links GPCR, N-acyl amide, bacterial genus and the site where these co-occur in the gastrointestinal tract (colored). Based on protein expression data (Human Protein Atlas) GPR119 is most highly expressed in the pancreas and duodenum, S1PR4 in the spleen and lymph node, G2A in the lymph node and appendix, PTGIR in the lung and appendix and PTGER4 in the bone marrow and small intestine. From gene expression data in the colon (GTEx dataset, N = 88 patient samples from small intestine, 345 patient samples from colon) GPR132, PTGER4, and PTGIR are all expressed alongside the N-acyl synthase genes known to encode metabolites that target these GPCR (Figure 1). In the gastrointestinal tract GPR119 and S1PR4 are most highly expressed in the small intestine where 16S studies have identified bacteria from the genera Gemella and Neisseria. All known reference genomes (NCBI) from these genera contain N-acyl synthase genes that are highly similar (blastN, e value 2e-132) to those we found to encode GPR119 or S1PR4 ligands.37,43,44 Cohen et al. Page 18 Nature. Author manuscript; available in PMC 2018 February 28. Author Manuscript Author Manuscript Author Manuscript Author Manuscript