Mouse Strains

The conditional LSL-Fbxw7^flox/flox mouse was a gift from Dr. Iannis Aifantis (New York University) [16]. Conditional LSL-Fbxw7^flox/flox; LSL-Kras^G12D/+; and P48-Cre strains were interbred to obtain LSL-Kras^G12D/+; LSL-Fbxw7^flox/flox; P48-Cre triple mutant animals (KFC). The mice were housed in specific pathogen-free facilities, and all the studies were conducted in compliance with the Institutional Committee on Use and Care of Animals guidelines. Genotyping was carried out by tail clipping from mice 2 weeks after birth. Tail specimens were incubated for 24 hours at 55°C in tail-lysis buffer containing proteinase K. NaCl was added, and cellular debris were pelleted by centrifugation. DNA was precipitated by the addition of isopropanol and washed with 70% ethanol. DNA pellets dissolved in water were used for polymerase chain reaction (PCR) analysis.