C57bl 6j apcmin

C57BL/6J (denoted as Apc^+/+ or wild-type) and C57BL/6J-Apc^Min/+ (denoted asApc^Min/+) mice were purchased from the Jackson Laboratory (Bar Harbor, ME) to develop breeding colonies that supplied all mice used in the study. Mice were maintained in micro isolator cages at 25 °C on a 12 h reverse light/dark cycle at the Case Western Reserve University (CWRU) Animal Resource Facility. Breeding was accomplished by placing one adult mutant (Apc^Min/+) male mouse with two WT adult female Apc^+/+ mice to generate mutant and WT littermates, which were subsequently distributed to experimental diets as previously described^{[2 (link)]}. Ear punch samples were taken from mice at 14 days of age for genotyping. Offspring were weaned and separated by sex at 21 days of age. Siblings were incubated together and fed a standard chow diet Prolab RMH 3000 (5P00; LabDiet; Brentwood, MO) with autoclaved water ad libitum. At 30 days of age, sibling male mice were separated by genotype, wild-type orApc^Min/+, distributed to an experimental diet (high-fat or low-fat) with autoclaved water, and then incubated in groups of two or three mice per cage until sacrifice after three days on experimental diet.

All of the mice were acclimated for two weeks in order to reduce stress from traveling. Carbon dioxide was used during euthanasia of the mice. This study was approved by the Institutional Animal Care and Use Committee (#202449) and Division of Laboratory Animal Resources at Stony Brook University. Three shipments of 10 four-week-old female C57BL/6J APC^Min/+ and 10 four-week-old female C57BL/6J WT mice were received from The Jackson Laboratory (Bar Harbor, ME) between June 2012 and May 2013. APC^Min/+ mice and WT mice were housed separately in groups of three to four in specific pathogen free (SPF) cages for two weeks prior to euthanization. All the experiments strictly followed guidelines from the Institutional Animal Care and Use Committee and Division of Laboratory Animal Resources at Stony Brook University.

C57BL/6 12.4KbVilCre transgenic mice were provided by D. Gumucio (University of Michigan, Ann Arbor, MI, USA). Lrp6^loxP/loxP and C57BL6/J-Apc^Min/+ mice were purchased from Jackson Laboratory. Lrp6^loxP/loxP; Villin-Cre mice (Lrp6^IEC-KO) were compared to Lrp6^loxP/+ or Lrp6^loxP/loxP control mice. For tumor initiation experiments, Lrp6^loxP/loxP; Villin-Cre; Apc^Min/+ mice were compared to Lrp6^loxP/+; Apc^Min/+ or Lrp6^loxP/loxP; Apc^Min/+ control mice. Genomic DNA was extracted from tissue by digestion in 25 mM NaOH/0.2 mM EDTA heated at 95 °C for 1 h followed by addition of an equal volume of 40 mM Tris-HCl (pH 5.5). Genotyping primer sequences and PCR conditions are available upon request. Experiments were approved by the Animal Research Ethics Committee of the Université de Sherbrooke (FMSS-399-18B) in accordance with the Canadian Council on Animal Care standards.