C57BL/6, CD45.1+, Cox10fl/fl, Rag1−/−, Foxp3RFP, Foxp3DTR-GFP, R26MYC, R26YFP reporter, and R26GFP reporter (a loxP site–flanked STOP cassette followed by the YFP- or GFP-encoding sequence inserted into the Rosa26 locus) mice were purchased from the Jackson laboratory. Foxp3YFP-Cre (25) and Foxp3Cre-ERT2 (42) mice were gifts from A. Rudensky. Mycfl/fl mice (9) were gifts from D.R. Green and F.W. Alt. Myc-GFP reporter mice (23) were gifts from B. Sleckman. Foxp3CreMycfl/fl mice were used at 2 to 3 weeks old, with age- and gender-matched control mice. Other mice were used at 8 to 10 weeks old, unless otherwise noted. Mixed BM chimeric mice were generated by adoptively transferring a 1:1 mix of CD45.1+ spike and CD45.2+ (Foxp3CreMycfl/+ or Foxp3CreMycfl/fl) T cell–depleted BM cells into sublethally irradiated (5.5 Gy) Rag1−/− mice, followed by at least 8 weeks of reconstitution. For Treg depletion experiments, Mycfl/+ and Mycfl/fl female mosaic mice (harboring a Foxp3DTR-GFP allele on one X chromosome and Foxp3Cre allele on the other X chromosome) were injected intraperitoneally with DT (50 μg kg−1; EMD Millipore) every other day for four total injections and then analyzed 3 days after the last injection. For tamoxifen administration, mice were injected intraperitoneally with tamoxifen (2 mg per mouse) in corn oil every other day for six total injections and then analyzed 3 weeks after the last injection. All mice were kept in a specific pathogen–free facility in the Animal Resource Center at St. Jude Children’s Research Hospital, and animal protocols were approved by the Institutional Animal Care and Use Committee.

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