Animal procedures complied with the guidelines of the European Union and protocols were approved by the local ethics committee of the Medicine Faculty, Université Libre de Bruxelles (CEBEA). Mouse strains were CD1 (Charles River), Cnx43-KI-CreER(T) (European Mouse Mutant Archive), Rosa26R-YFP and Rosa26R-Tomato (both from The Jackson Laboratory). Lgr5-DTR-EGFP knock-in mice were kindly provided by Genentech (Tian et al., 2011 (link)). The day the vaginal plug was observed was considered as E0.5.
For lineage-tracing experiments, tamoxifen (Sigma-Aldrich) was dissolved in sunflower oil (Sigma-Aldrich)/ethanol mixture (9:1) at 10 mg/ml. Pregnant females were injected intraperitoneally at 0.1 mg/g body weight. For specific cell ablation in Lgr5-DTR mice, diphtheria toxin (Sigma-Aldrich) was injected intraperitoneally (50 µg/kg) into 8-week-old mice. Control mice were injected with sterile PBS solution. Eight-week-old CD1 mice were injected subcutaneously with 300 µg/kg indomethacin (dissolved in 0.6 M NaHCO3 buffer pH 8.5 and 5% DMSO) or with the buffer as control.
For lineage-tracing experiments, tamoxifen (Sigma-Aldrich) was dissolved in sunflower oil (Sigma-Aldrich)/ethanol mixture (9:1) at 10 mg/ml. Pregnant females were injected intraperitoneally at 0.1 mg/g body weight. For specific cell ablation in Lgr5-DTR mice, diphtheria toxin (Sigma-Aldrich) was injected intraperitoneally (50 µg/kg) into 8-week-old mice. Control mice were injected with sterile PBS solution. Eight-week-old CD1 mice were injected subcutaneously with 300 µg/kg indomethacin (dissolved in 0.6 M NaHCO3 buffer pH 8.5 and 5% DMSO) or with the buffer as control.
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