The OECD Principles of GLP was developed specifically for the conduct of studies which produce data for regulatory decisions. The experiment on GLU-VENUS mice was approved by local ethics committees and conformed to United Kingdom Home Office regulations, the experiment on Tgr5−/− mice were approved by the local animal experimentation committee of the Canton de Vaud (license no. 2614) and the experiment on GF/CONV-R mice was performed with protocols approved by the University of Gothenburg Animal Studies Committee. After 30 min glucose deprivation in DMEM No Glc medium, a 1 h-GLP-1 secretion test in response to DMEM without glucose, to DMEM with glucose (5.5 mmol l−1) or in response to DMEM with glucose (5.6 mmol l−1) plus phloretin (0.5 mmol l−1) was performed at 37 °C in DMEM plus 1% DPP-4 inhibitor (Ile-Pro-Ile, Sigma-Aldrich). After permeabilization during 10 min (Tris/NaCl/0.1% TRITON X100), unspecific protein binding sites were masked by Dako Protein Block (DAKO) during 2 h at room temperature. The remaining tissue was washed in cold PBS and frozen at −80 °C in NaOH 2 N. Protein content was assessed according to BCA’s method (Thermoscientific). Cells were lysed in NaOH (0.8 mol l−1) under agitation and total protein content was determined using the BCA Protein Assay Kit (Pierce).

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