Bionic buffer

Culture media and supplements, collagenase type XI, histopaque-1077, DMSO, EDTA, IBMX, carbachol, clonidine, LiCl, exendin-4, forskolin, agarose, bionic buffer and BSA were obtained from Sigma-Aldrich (Dorset, UK). DNeasy Blood and Tissue, RNeasy Mini and QuantiTect SYBR Green PCR kits and qPCR primers for mouse and human CB₁ (CNR1), GPR119, GPR18, GPR92 (LPAR5), delta-opioid receptor (OPRD1), transient receptor potential cation channel subfamily V member 1 (TRPV1), GPR3, GPR6, GPR12, and ACTB were from Qiagen (Manchester, UK). PCR primers for Gpr55 genotyping were from Eurofins Genomics (Wolverhampton, UK). SR141716A was from Tocris Bioscience (Abingdon, UK). AM251 and rabbit anti-Ki67 primary antibody were from Abcam (Cambridge, UK). cAMP HiRange and IP-one (IP₁) assays were from Cisbio (Codolet, France). TaqMan RT-PCR kit, 100 base pairs (bp) DNA ladder, SYBR® DNA gel stain, HEPES, HBSS and DAPI were from Thermo Fisher Scientific (Paisley, UK). Caspase-Glo 3/7 and GoTaq^® G2 Green Master Mix were from Promega (Southampton, UK). Recombinant TNFα, IFNγ and IL-1β were from PeproTech EC (London, UK). Guinea pig anti-insulin was obtained from Dako (Cambridge, UK). AlexaFluor 488- and AlexaFluor 594-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (Newmarket, UK).

Thermo Scientific DreamTaq PCR Master Mix (2X) (K1071) was used for all PCR reactions. PCR products ran on a 0.7–2% agarose gel in 1X Bionic buffer (Sigma-Aldrich) to verify that the PCR reaction progressed as expected. The percentage of agarose was determined by how large a PCR product was expected. Monarch® DNA Gel Extraction Kit Protocol (NEB #T1020) was used to extract DNA from a gel slice and purify the PCR product for Sanger sequencing. Sanger sequencing was performed by Eurofins Genomics.

Culture media and supplements, collagenase type XI, histopaque-1077, DMSO, EDTA, IBMX, carbachol, clonidine, Accutase, Fura-2 AM, tolbutamide, LiCl, exendin-4, forskolin, agarose, bionic buffer, and BSA were obtained from Sigma-Aldrich (Dorset, UK). JTE 907 was from Tocris Bioscience (Abingdon, UK). Rabbit anti-Ki67 primary antibody was from Abcam (Cambridge, UK). cAMP HiRange and IP-one (IP₁) assays were from Cisbio (Codolet, France). HEPES, HBSS, and DAPI were from Thermo Fisher Scientific (Paisley, UK). Caspase-Glo 3/7 was from Promega (Southampton, UK). Recombinant TNFα, IFNγ, and IL-1β were from PeproTech EC (London, UK). Guinea pig anti-insulin was purchased from Dako (Cambridge, UK). Alexa Fluor 488- and Alexa Fluor 594-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (Newmarket, UK).