Rm3 e diet

All animal work was performed under the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012 following ethical review by the University of Cambridge Animal Welfare and Ethical Review Body (AWERB). All investigators understood and worked by the ethical principles and standards discussed by Grundy (2015 (link)). Mice were housed in M3 conventional cages (NKP, UK), at 55% humidity and 21°C, with a 12 h light cycle. Mice were fed RM3(E) diet (Special Diet Services) ad libitum from weaning. C57Bl/6 mice were purchased from Charles River Laboratories and bred in‐house. Pregnant females were identified on the basis of a vaginal plug. Noon of the day the vaginal plug was identified was dated as E0.5.

Animals were maintained on an outbred background, housed in a standard facility and all experimental procedures were performed in accordance with institutional animal care and ARRIVE guidelines in compliance with UK Home Office regulations. In brief, mice were maintained in conventional open top cages on dust free bedding (IPS Ltd) under a 12 hr light cycle, with RM3(E) diet (Special Diet Services UK) provided for nutritional support. To enrich the environment, sunflower seeds (at weaning only, LBS Ltd), nestlets (IPS Ltd), disposable envirotubes (IPS Ltd) and small chewsticks (Labdiet–IPS Ltd) were provided. Mice carrying the targeted Lect2 allele were kindly supplied by Dr Satoshi Yamagoe [5 (link)]. Experimental mice were genotyped as previously described for the targeted Lect2 allele [5 (link)], Apc allele [10 (link)], the Apc^Min/+ allele [39 (link)], the Rosa26R allele [40 (link)] and the AhCre transgene [11 (link)]. Cre activity was induced in control and experimental mice by 3 consecutive intraperitoneal (i.p.) injections of 80 mg/kg β-naphthoflavone (Sigma, UK) in 24 h. Mixed sex control (litter mates) and experimental mice were used for timepoint (N > 4) or survival (N > 15) experiments. Prior to proliferation analysis selected animals were injected with 100 μg/kg Bromo-deoxyuridine (Sigma, UK) and culled at indicated time points after labelling.