B6.129p2 lgr5tm1 cre ert2 cle j

C57BL/6 mice were bred in Tierforschungszentrum (TFZ), Ulm University. Tg(Vil-cre/ERT2)23Syr, Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}/J, and B6.129P2-Lgr5^{tm1(cre/ERT2)Cle}/J were purchased from the Jackson Laboratory. Tg (Vil1-cre/ERT2) mice were crossed with Confetti; R26 to get vil1-cre; R26 Confetti mice and Lgr5EGFP-ires-creERT2 were crossed with Confetti; R26 to get Lgr5e-creERT2; R26 Confetti. All transgenic mice were backcrossed to C57BL/6 mice. To initiate the confetti fluorophores either in the whole intestinal epithelial cells or specifically in the Lgr5 cells, vil1-cre; R26 Confetti or Lgr5e-creERT2; R26 Confetti mice, respectively, were injected intraperitoneal with a single dose (5 mg) of tamoxifen (Sigma-Aldrich). All mice were housed in the animal barrier facility under pathogen-free conditions at the Ulm University. All mouse experiments were performed in compliance with the German Law for Welfare of Laboratory Animals and were approved by the Institutional Review Board of the Ulm University and by the Regierungspraesidium Tuebingen (state government of Baden-Württemberg), protocol number: 35/9185.81-3/1407. Housing conditions: a temperature range of 22°C ± 1°C, a relative humidity of 55% ± 10%, an air change rate of 15 times, and a light/dark change of 12/12 h. Nutrition: ssniff M-Z autoclavable complete for mice breeding (#V1124-3).

C57BL/6J, B6.129P2-Lgr5^{tm1(cre/ERT2)Cle}/J, and B6.129-Cdh1^tm2/Kem/J mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA). BALB/c mice were purchased from Orient Bio Inc. (Seongnam, Rep. of Korea). All mice were maintained at the Korea Research Institute of Bioscience and Biotechnology (Daejeon, Rep. of Korea). Lgr5-CreERT2;Cdh1^fl/fl mice were generated by cross-breeding B6.129P2-Lgr5^{tm1(cre/ERT2)Cle}/J and B6.129-Cdh1^tm2/Kem/J mice. All mice were housed under a constant 12-h light/dark cycle at a suitable temperature (22 ± 2 °C) and humidity (typically 55 ± 5%) with a standard chow diet (Teklad Global 18% Protein Rodent Diet 2018S; 18.6% crude protein, 6.2% fat, 44.2% available carbohydrates; Envigo, Indianapolis, IN, USA) and water provided ad libitum in a specific pathogen-free facility. Mice between 2 and 4 months of age were considered young and mice over 22 months of age were considered old. 100-week-old male C57BL/6 mice were treated orally with vehicle or Amuc_1409^* (4.5 μg per mouse) daily for 15 weeks. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Korea Research Institute of Bioscience and Biotechnology (KRIBB-AEC-22244, KRIBB-AEC-22193) and were performed in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health.

Lgr5-EGFP-IRES-Cre^ERT2 knock-in mice (B6.129P2-Lgr5^{tm1(cre/ERT2)Cle}/J; JAX mice #008875) and ROSA26-LSL-LacZ mice (B6.129S4-Gt (ROSA)26Sor^tm1Sor/J; JAX mice #003474) were purchased from the Jackson Laboratory (ME, USA). The mice were bred in a conventional clean-room facility at the Central Research Institute of Electric Power Industry (CRIEPI) under conditions of controlled temperature (24 ± 2°C) and humidity (45% ± 5%), with a 12-h light–dark cycle and ad libitum access to γ-sterilized food (CLEA Japan, Tokyo, Japan) and filter-sterilized deionized water. All animal experiments were approved by the Animal Research and Ethics Committee at the CRIEPI and were performed in accordance with the guidelines for animal care in Japan.