Wnt1-Cre;R26RTomato mice were killed by cervical dislocation followed by dissection of small intestine. During all steps the tissue was kept in aCSF (in mM: 118 NaCl, 4.6 KCl, 1.3 NaH2PO4, 25 NaHCO3, 20 glucose, 7 mM CaCl2 and MgSO4) equilibrated in 95% O2 5% CO2 for 30 min before use and held on ice. The small intestines of male and female (P21) mice were cut in 5cm pieces and flushed clean with ice-cold aCSF using a blunt 20G needle attached to a 20ml syringe. The mesentery was removed, the pieces opened lengthwise along the mesenteric border and pinned with the mucosa side down on a Sylgaard (Dow Corning) covered dissection dish. The outer smooth muscle layers, containing the myenteric plexus were peeled off from the submucosa using forceps. The tissue was digested in 1,5 mg/ml Liberase (Grundmann et al., 2015), 0.1 mg/ml DNaseI and 1xAntibiotic-Antimycotic (ThermoFisher) in aCSF at 37°C for 1h, with shaking of the tube every 15 min. The cells were gathered by centrifugation at 356 g for 5min followed by incubation in TrypLE for 30 min. The
