Vismodegib (V-4050, LC Laboratories) was administered i.p.55 (link)56 (link) in dimethyl sulfoxide twice daily at 25 mg kg−1 body weight for 6 days per week.
For intracolonic administration, 4OH-Tam (H6278; Sigma-Aldrich) was dissolved in 99.5% ethanol, vortexed, further diluted in corn oil (S5007, Sigma-Aldrich) to reach the working concentration and sonicated for 30 min at 37 °C in a water bath. The solution was kept at 37 °C in darkness until it was instilled intraluminally at 1 mg 25 g−1 bodyweight. Instillation was performed with US guidance under isoflurane anaesthesia (described below). For visualization of the instilled substance with microcomputed tomography (μCT) imaging, 0.1 mg ml−1 Barium Sulphate (Barium Sulphate (#11381499) 97%, Alfa Aesar) was added to the Tam/oil suspension. The μCT was performed under isoflurane anaesthesia (see below) using a Quantum FX μCT imaging system (Caliper Life Sciences, Perkin Elmer, Inc.) a few minutes after administration of the Tam/oil/barium mixture.
For systemic labelling experiments in Col1a2CreER;R26-LSL-tdTomato and Gli1CreERT2;R26-LSL-tdTomato mice, 5 mg of Tam was dissolved in corn oil using sonication at 37 °C at 20 mg ml−1 and injected once i.p.
For intracolonic administration, 4OH-Tam (H6278; Sigma-Aldrich) was dissolved in 99.5% ethanol, vortexed, further diluted in corn oil (S5007, Sigma-Aldrich) to reach the working concentration and sonicated for 30 min at 37 °C in a water bath. The solution was kept at 37 °C in darkness until it was instilled intraluminally at 1 mg 25 g−1 bodyweight. Instillation was performed with US guidance under isoflurane anaesthesia (described below). For visualization of the instilled substance with microcomputed tomography (μCT) imaging, 0.1 mg ml−1 Barium Sulphate (Barium Sulphate (#11381499) 97%, Alfa Aesar) was added to the Tam/oil suspension. The μCT was performed under isoflurane anaesthesia (see below) using a Quantum FX μCT imaging system (Caliper Life Sciences, Perkin Elmer, Inc.) a few minutes after administration of the Tam/oil/barium mixture.
For systemic labelling experiments in Col1a2CreER;R26-LSL-tdTomato and Gli1CreERT2;R26-LSL-tdTomato mice, 5 mg of Tam was dissolved in corn oil using sonication at 37 °C at 20 mg ml−1 and injected once i.p.