Rag1 deficient mice

Foxp3^ΔEGFP (FoxP3- deficient) and Foxp3^EGFP mice were the gift of Dr. Talal Chatila (Harvard University). CD4-deficient and IL-10-deficient mice on a C57BL/6 genetic background, wild-type C57BL/6, and Rag1-deficient mice were purchased from Jackson Laboratory. Experimental and control mice were 8- to 10-week-old. Mice were maintained in the Biological Resource Center at the Medical College of Wisconsin. All animal protocols were approved by the MCW Institutional Animal Care and Use Committee.

C57BL/6 mice were obtained from the Animal Production Facility of the National Cancer Institute –Frederick Cancer Research and Development Center (Frederick, MD) and Rag1-deficient mice, B6 background, from The Jackson Laboratory (Bar Harbor, ME). The mice carrying the Cre-dependent conditional alleles for Crk and CrkL, generously provided by Dr. A Imamoto, were generated in accordance to animal protocols approved by the IACUC of the University of Chicago. The Crk conditional allele was generated by inserting two loxP sites upstream and downstream of Exon 1 by homologous recombination in a line of ES cells with a B6 inbred background. A mouse strain carrying a CrkL allele, CrkL^{tm1a(EUCOMM)Hmgu}, was generated using ES cells in a B6-inbred background, obtained from EUCOMM/Helmholtz Zentrum Muenchen [38 (link),39 (link)]. To make it a Cre-dependent conditional allele, the lacZ and neo cassettes were removed by genetic cross with a FLP strain to derive a conditional allele, CrkL^{tm1c(EUCOMM)Hmgu}, leaving only one FRT and one loxP sites upstream of Exon 2 and one loxP site downstream of Exon 2. For the current study, the Crk and CrkL conditional strains were intercrossed to obtain compound homozygotes. Animal care was provided in accordance with procedures outlined in the “Guide for Care and Use of Laboratory animals” (National Institute of Health, Bethesda, MD, 1996).

Slc1a5^−/− mice (in C57BL/6 × 129/sv genetic background) were generated by a conventional gene-targeting strategy, in which coding exons 2 and 4 were replaced with a lacZ–neomycin-resistance cassette (Taconic, Figure S1). Heterozygous Slc1a5^+/– mice were bred to generate age- and sex-matched homozygous ASCT2-ablated (Slc1a5^−/−) and wild-type (Slc1a5^+/+) mice, which were used in the experiments at the indicated ages. Genotyping PCR and RT-PCR primers are listed in Supplementary Table1. The TCR transgenic OT-II mice and the lymphocyte-deficient Rag1^−/− deficient mice were from Jackson Laboratory. Ikbkb-floxed mice, provided by Dr. Manolis Pasparakis (University of Cologne), were crossed with Cd4-cre mice (Jackson Laboratory) to produce T cell-conditional Ikbkb^−/− mice. Card11^−/− mice (C57BL/6 × 129/sv genetic background) were provided by Dr. Josef Penninger (Austria Academy of Sciences), Bcl10^−/− mice were provided by Dr. Stephan Morris (St Jude Children’s Research Hospital), and the Malt1^−/− mice were provided by Dr. Vishva Dixit (Genentech). Mice were maintained in specific pathogen-free facility of The University of Texas MD Anderson Cancer Center, and all animal experiments were conducted in accordance with protocols approved by the Institutional Animal Care and Use Committee.

Kras^fl/fl mice were crossed with VavCre transgenic mice and the mouse line was maintained on C57BL/6 genetic background (> N10) (22 (link)). Experimental VavCreKras^fl/fl and control VavCreKras^fl/+ mice were 8-12 weeks old. BALB/c (H-2^d) and Rag1-deficient mice were from Jackson Laboratory. The Medical College of Wisconsin Institute Animal Care and Use Committee approved the animal protocols.