C57bl 6 tg tcratcrb 1100mjb j ot 1 mice

All mice were housed at University of Virginia specific pathogen-free facilities with a 12 hr light/dark cycle. C57BL/6 J (#000664), CBA/J (#000656), B6.Cg-Tg(Itgax-Venus)1Mnz/J (CD11c^YFP mice, #008829), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I mice, #003831), and C57BL/6-Tg(Nr4a1-EGFP/cre)820Khog/J (Nr4a1^GFP mice, #016617) strains were originally purchased from the Jackson Laboratory and then maintained within our animal facility. B6.SJL-Ptprc^aPepc^b/BoyCrCrl (CD45.1 congenic mice, #564) and Swiss Webster (#024) strains were purchased from Charles River Laboratories. To generate OT-I (CD45.1 congenic) mice, OT-I and CD45.1 congenic strains were cross-bred within our facility. OT-I (CD45.1 congenic) mice were then crossed to Nr4a1^GFP mice to generate OT-I/Nr4a1^GFP (CD45.1 congenic) mice for use in adoptive transfer studies. For experiments reported in this study, age-matched young adult female mice (7–10 weeks of age) were used. All experiments were approved by the Institutional Animal Care and Use Committee at the University of Virginia under protocol number 3968.

All animals were maintained in a specific pathogen–free facility. Whsc1-floxed mice were generated by the Beijing Biocytogen Company as previously described (25 (link)). C57BL/6-Tg (TcraTcrb) 1100Mjb/J (OT-I) mice, Apc^min/+ mice, and Villin^Cre/+ mice were purchased from The Jackson Laboratory. All mice were backcrossed with C57BL/6 mice for at least 7 generations. BALB/c, C57BL/6, Rag1^–/–, or NSG male mice aged 4 to 8 weeks were injected subcutaneously with 1 × 10⁶ CT26, MC38, CRC610301, or CRC541051 cells. For cecal injections, C57BL/6 and Rag1^–/– male mice aged 4 to 8 weeks were injected with 5 × 10⁵ KAP cells (derived from Villin^Cre/+Kras^G12DApc^min/+Trp53^fl/fl mice). An isotype or anti-CD8 antibody (10 mg/kg, Bio X Cell, BP0117) was intravenously administered every 3 days. For anti–PD-1 treatment, 10 mg/kg anti–PD-1 (Bio X Cell, BE0273) or an isotype antibody was intravenously administered every 3 days after 8 to 10 days of tumor cell implantations. For IFN-γ treatment, 25 μg/kg IFN-γ (GenScript, Z02915) was intravenously administered over 3 consecutive days after 30 days of tumor cell implantations.