All experiments were performed according to US National Institutes of Health guidelines and were approved by the IACUC of The Cincinnati Children’s Hospital. C57BL/6J mice were obtained from Jackson. Deletion of Gimap5 in Gimap5flox/floxCd4cre-ert2 mice was induced by the administration of tamoxifen chow (40mg/kg body weight; Harlan Laboratories Teklad Diets). Gimap5sph/sph mice were generated as previously described (6) and bred in-house in the vivarium of Cincinnati Children’s Hospital. All mice were maintained under specific pathogen-free conditions.
All antibodies used for flow cytometry were purchased from eBioscience or Biolegend unless otherwise noted. Purified α-mouse-CD3 (17A2) and α-mouse-CD28 (37.51) antibodies (Biolegend) were used for mouse T cell activation. A fixable viability stain was purchased from eBioscience. 7-AAD was purchased from BD. PMA, ionomycin, and Brefeldin A were obtained from Sigma. αIFNγ (XMG1.2) and mIL-23 were obtained from eBioscience. TGFβ1, mIL-4, and hIL-2 were purchased from Miltenyi Biotech. αIL-4, IL-6, and mIL-1β, mIL-12, and TGFβ3 were obtained from PeproTech.
All antibodies used for flow cytometry were purchased from eBioscience or Biolegend unless otherwise noted. Purified α-mouse-CD3 (17A2) and α-mouse-CD28 (37.51) antibodies (Biolegend) were used for mouse T cell activation. A fixable viability stain was purchased from eBioscience. 7-AAD was purchased from BD. PMA, ionomycin, and Brefeldin A were obtained from Sigma. αIFNγ (XMG1.2) and mIL-23 were obtained from eBioscience. TGFβ1, mIL-4, and hIL-2 were purchased from Miltenyi Biotech. αIL-4, IL-6, and mIL-1β, mIL-12, and TGFβ3 were obtained from PeproTech.