Ptentm1hwu

Mice were housed in AAALAC-accredited vivarium. Room temperature was maintained at 22 ± 3 °C and the humidity at 55 ± 15%. Animals were kept at a 12 h/12 h dark/light cycle. The C57BL/6 and SCID/Beige mice were purchased from Charles River (Wilmington, MA). Nude mice, Col1a2-CreER^T2, C57BL/6-Tg(TRAMP)8247Ng/J, and B6.Cg-Gt(ROSA)26Sor^{tm3(CAG-EYFP)Hze}/J, Pten^tm1Hwu, and R26-LSL-Kras^G12D mice were purchased from the Jackson Laboratory (Bar Harbor, ME). The ARR2Pb-Cre mice were from Dr. Fen Wang at the Institute of Biosciences and Technology at Texas A&M. The R26-LSL-Foxf2 mice were generated at the Baylor College of Medicine. The Foxf2 null mouse embryos were generously provided by Dr. Rulang Jia at Cincinnati Children’s Hospital. All mice were on the C57BL/6 background. Mice were genotyped by polymerase chain reaction using mouse genomic DNA from tail biopsy specimens. The sequences of genotyping primers and the expected band sizes for PCR are listed in Supplementary Table 2. PCR products were separated electophoretically on 1% agarose gels and visualized via ethidium bromide under UV light.

All animal study protocols were approved by the University of Medicine and Dentistry of New Jersey Institutional Animal Care and Use Committee review board. Conditional alleles for Pten (Pten^Tm1Hwu) (Lesche et al.^{19 (link)}) and p53 (Trp53^Tm1Brn, Jonkers et al.^{18 (link)}) mice were purchased from Jackson Laboratories, Bar Harbor, ME, USA. In all, 8–10-week old mice were used for injection of an adenovirus expressing cre recombinase (Ad5CMVCre, University of Iowa's Vector Core Facility (http://www.uiowa.edu/~gene) into adipose tissue. Concentrated virus (25 μl; 4 X10¹¹PFU/ml) was mixed with 20 μl of Dulbecco's modified eagle medium (D-MEM). Mice were monitored for tumor development by palpitation at weekly intervals. Kaplan–Meier analysis was generated using GraphPad software.

B6.129-Pten^tm1Rps (Pten^+/−)55 (link) were obtained from the repository at the National Cancer Institute at Frederick, where they were already backcrossed onto a congenic C57BL/6J background by the Donating Investigator. The line has been maintained by backcrossing to C57BL/6J mice for more than 10 generations. All other mouse lines were obtained from the Jackson Laboratory: Etv1^{tm1.1 (cre/ERT2)Zjh}(Etv1-CreERT2, stock number 013048), Gt (ROSA)26Sor^{tm14 (CAG−tdTomato)Hze} (Ai14, stock number 007914), Rptor^tm1.1Dmsa(Rptor^loxp/loxp, stock number 013188), Emx1^{tm1 (cre)Krj} (Emx1-Cre⁺, stock number 005628), Pten^tm1Hwu(Pten^loxP/loxP, stock number 006440), and Fmr1^tm1Cgr (Fmr1^−/y, stock number 003025). To obtain Emx1-Cre⁺; Rptor^loxp/+; Pten^+/−mice, Rptor^loxp/loxp; Pten^+/− male mice were mated with Emx1-Cre⁺ female mice. Rptor^loxp/+ mice were used as controls. All animal experiments were conducted in accordance with NIH and AAALAC guidelines and were approved by The Scripps Research Institute's Institutional Animal Care and Use Committee.