All animals were housed in a temperature-controlled environment under a 12h light-dark cycle with free access to water and a standard rodent chow diet (Lab Diet 5001), unless otherwise specified. All animal studies were approved by the Salk Institute Institutional Animal Care and Use Committee. TORC1 mutant mice were generated by insertional mutagenesis. Mouse embryonic stem (ES) cells containing an insertional gene-trap in the CRTC1 locus (XK522; 129/Ola mouse strain) were obtained from BayGenomics 42,43. ES cells were injected into C57BL/6 blastocysts to generate chimeric mice, which were backcrossed with C57BL/6 mice (Harlan). Prior to intercrossing, heterozygous mice were backcrossed with C57BL/6 mice for three successive generations. The heterozygous progeny were intercrossed to obtain homozygous, heterozygous and wild-type littermate animals.
