Ro 64 5229

Recombinant rat NL1 (ecto-NL1) was obtained from R&D Systems (Cat. No. 4340-NL-050, Abingdon, UK). Expression vectors that encoded N-terminally HA-tagged mouse NL1-AB, NL1-A, NL1-B and NL1-0 (Addgene plasmid no. 15262, 15227, 15261 and 15260, respectively; Chih et al., 2006 (link)) and NL1 without esterase-homology domain (NL1∆ACh; Scheiffele et al., 2000 (link)) were gifts from Peter Scheiffele. In the NL1∆ACh deletion mutant, the amino acids 48–630 of murine NL1AB were excised and the HA epitope is followed by the amino acids GVPHLHN. Expression vector that encoded mouse C-terminally mGluR2-green fluorescent protein (GFP) was purchased from Origin (Rockville, MD, USA). Mouse mGluR2 was subcloned into a pcDNA5FRT vector (Life Technologies, Waltham, MA, USA) to create an expression vector that encoded C-terminally strep-tagged mGluR2. The pharmacological inhibitor of mGluR2, RO 64-5229, was purchased from Tocris (Bristol, UK).

All chemicals were purchased from Sigma-Aldrich, Merck and Roth unless otherwise noted. n-dodecyl-β-D-maltopyranoside (DDM), lauryl maltose neopentyl glycol (LMNG), and cholesteryl hemisuccinate (CHS) tris salt were purchased from Anatrace (through CliniSciences, France). Glyco-diosgenin (GDN) was purchased from Avanti Polar Lipids through Merck. SNAP-Lumi4-Tb, SNAP-green, SNAP-Cy3B, and SNAP-d2 were obtained from Cisbio Bioassays (Codolet, France). DCG-IV, LY341495, LY379268, LY354740, LCCG-I, BINA hydrochloride, and Ro64-5229 were purchased from Tocris Bioscience (Bristol, UK).

LY 379268, a highly selective group II mGlu receptor agonist (Tocris Bioscience, Rennes, France), was dissolved in sterile water. LY 2389575 hydrochloride, a selective negative allosteric modulator of mGlu3 receptors (Tocris), and Ro 64-5229, a selective, non-competitive mGlu2 receptor antagonist (Tocris), were dissolved in DMSO.