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. 2023 Feb 8;220(4):e20221288. doi: 10.1084/jem.20221288

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Figure 7. — CTCF has an early impact on T_EFF and T_MP gene expression in antigen-experienced CD8⁺ T cells. (A) Tracks of CTCF CUT&RUN, ATAC-seq, and Tcf1 ChIP-seq at the Tcf7 and Id3 gene loci linked to the incongruous sites (denoted with blue bars). The green bars denote ChrAcc⁻ constitutive CTCF binding sites that function as boundary anchors for insulated neighborhoods. (B–G) Detection of T_EFF- and T_MP-characteristic proteins in CTV-labeled WT or Ctcf⁻ CD45.2⁺GFP⁺CD8⁺ T cells within 60 h after in vivo activation by LCMV-Arm infection. B and C, intracellular detection of granzyme B; D and E, intracellular detection of GP33-stimulated IFN-γ production; F and G, intranuclear detection of Tcf1. In B, D, and F, representative dot plots showing percentage of actively dividing cells that expressed proteins of interest; in C, E, and G, half-stacked histographs showing gMFI of the proteins. (H) Detection of active caspase3/7 in CTV-labeled WT or Ctcf^−/− CD8⁺ T cells within 60 h after in vivo activation. Cumulative data in B–H are means ± SD (right) from at least two independent experiments. *, P < 0.05; ***, P < 0.001 by two-tailed Student’s t test. gMFI, geometric mean fluorescence intensity.