C57bl 6 tg tcratcrb 1100mjb j ot

Four to six week old C57BL/6, Rag1^−/−, K-ras^LSL-G12D, Trp53^L/L and C57BL/6-Tg (TcraTcrb)1100Mjb/J OT-I mice were purchased from Jackson labs. K-ras^LSL-G12D and Trp53^L/L mice were crossed to obtain K-ras^LSL-G12D/+; Trp53^L/L mice (DuPage et al., 2009 (link)). OT-I Tg mice were crossed to Foxp3^{Cre-YFP.Thy1.1} mice. Ebi3^Tom and Ebi3^{Tom.L/L.Thy1.1} mice were generated in our laboratory as described in supplementary information (Figures S5 and S3). Foxp3^Cre-YFP (Rubtsov et al., 2008 (link)) and Foxp3^DTR-gfp (Fontenot et al., 2005 (link)) mice were obtained from A.Y. Rudensky and crossed to Ebi3^Tom mice. Nur77^GFP mice were obtained from K.A. Hogquist (Moran et al., 2011 (link)) and crossed to Foxp3^Cre-YFP.Ebi3^Tom mice. Foxp3^DTR-gfp mice were treated with diphtheria toxin (DT) (Sigma, St. Louis, Missouri) as described (Kim et al., 2007 (link)). Mouse experiments were carried out in AAALAC-accredited Helicobacter–, MNV–, and specific pathogen-free facilities at St. Jude Children’s Research Hospital and University of Pittsburgh in accordance with IACUC guidelines.

Six- to ten-week-old C57BL/6, B6.SJL-ptprc^a (CD45.1⁺), B6.129S2-Cd4^tm1Mak/J (CD4 KO), B6.129S2-H2^dlAb1-Ea/J (MHC II KO), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I), and B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II) mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Thymectomized animals underwent adult thymectomy at the Jackson Laboratory (Bar Harbor, ME). E1/E3 deleted Ad5-SIINFEKL-Luc, Ad5-OVA, and Ad5HVR48(1–7)-Gag have been previously described (35 (link)–38 (link)). Mice were immunized intramuscularly with 10⁹ viral particles of the indicated vector in the quadriceps using 100 µl divided equally between the two legs. NYVAC-Env has been previously described (39 (link)) and was used at 10⁶ pfu per mouse. All animal experiments were performed in accordance with the Institutional Animal Care and Use Committee guidelines of Beth Israel Deaconess Medical Center.

C57BL/6NTac mice were purchased from Taconic Farms. B6.SJL-Ptprca/BoyAiTac (CD45.1), OT-I CD45.1 RAG^−/− (OT-I) were obtained through the NIAID Taconic exchange program. C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I) and B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J (mTomato) mice were purchased from the Jackson Laboratories and crossed to generate mTomato OT-I mice. C57BL/6-Tg(Adipoq-cre/ERT2)1Soff/J and B6.129P2-Gt(ROSA)26Sor^tm1(DTA)Lky/J were purchased from Jackson laboratories and crossed to generate Adipoq-Cre^ERT2 x Rosa26-DTA mice. B6Cg.Thy1a/Cy Tg(TcraTcrb)8Rest/J (pmel) were from the laboratory of N.P.R. All experiments involved female mice 6–23 weeks of age. Mice were bred and maintained under specific pathogen-free conditions at an American Association for the Accreditation of Laboratory Animal Care (AAALAC)-accredited animal facility at the NIAID and housed in accordance with the procedures outlined in the Guide for the Care and Use of Laboratory Animals. Cxcr4^fl/fl x UBC-CRE^ERT2 and S1pr1^fl/fl x UBC-CRE^ERT2 mice were housed at the Skirball institute animal facility. Experiments were performed under an animal study proposal (LISB-18E, 19E and 20E) approved by the NIAID Animal Care and Use Committee.