Fig 1: TET2 promotes inflammatory cytokine-mediated demethylation of murine Ifng during effector CD8 T cell differentiation(A) Experimental setup showing adoptive transfer of CD45.1 OT-1 CD8 T cells into WT C57BL/6 CD45.2 mice. One day later, the mice were either infected with LM-OVA or co-infected with LM-OVA and LCMV Armstrong. At the effector (8 days) and memory (>2 months) stages, CD45.1 OT1 CD8 T cells were FACS purified for phenotypic and DNA methylation analysis.(B) Representative and summary graphs of IFNg CNS bisulfite sequencing methylation profiles of naive CD8 T cells and LM-specific and LCMV-specific effector CD8 T cells at day 8 postinfection. The bottom bar graphs show the percentage of CpG methylation (mean ± SEM) at each individual CpG site of the IFNg locus. n = 3 for LM and n = 2 for LM+LCMV summary graphs.(C) Experimental setup showing infection of WT C57BL/6 or TET2cKO mice with LCMV Armstrong. At the effector (8 days) stage, LCMV-specific effector CD8 T cells were FACS purified for DNA methylation analysis.(D) Representative and summary bisulfite sequencing methylation profiles of the Ifng 3' CNS in LCMV-specific effector CD8 T cells from WT or TET2cKO mice. Error bars are defined based on the means ± SEM.(E) Experimental setup showing adoptive transfer of Thy1.1+ P14 CD8 T cells into Thy1.2+ WT C57BL/6 or TET2cKO mice. One day later, the mice were infected with LCMV Armstrong. At the effector (8 day) stages, endogenous and adoptively transferred GP33-specific CD8 T cells were phenotypically characterized and FACS purified for DNA methylation analysis.(F) Representative FACS analysis and summary graph characterizing the terminal effector (KLRG1+ CD127-) and memory precursor (KLRG1- CD127+) phenotype of either endogenous or adoptively transferred LCMV-specific CD8 T cells isolated from WT C57BL/6 or Tet2cKO mice. n = 4 for Tet2cKO and n = 4 for WT C57BL/6 summary graphs. ****p < 0.0001.(G) Targeted methylation profiling of the IFNg 3' CNS in naive, endogenous, and WT P14s CD8 Tcells isolated from WT and TET2cKO mice at the effector time point.