T _EFF -acquired CTCF binding activates effector transcriptional program and is partly dependent on Tbet and Runx3. (A) Tracks of CTCF CUT&RUN, ATAC-seq, and Tcf1 ChIP-seq at the genes in the cytotoxic program. Orange bars denote T_EFF-acquired CTCF binding sites linked to CTCF-dependent ChrAcc in T_EFF cells, and bars with dotted lines denote CTCF binding sites that depended on Tbet and Runx3 (TRKO, Tbx21^−/−Runx3^−/−). (B) Detection of Tbet and Runx motifs in the congruous CTCF sites based on HOMER analysis. Values are motif⁺ site numbers and those in parentheses denote detection frequency. (C) PCA of the CTCF binding profiles of WT and Tbx21^−/−Runx3^−/− early T_EFF cells, where normalized CTCF CUT&RUN signals on merged peaks were used as input data. (D) Top motifs in Tbet/Runx3-dependent CTCF sites (5,321 sites as defined in Fig. 6 H) based on HOMER analysis. (E) Distribution of Tbet/Runx3-dependent CTCF sites among T_EFF-acquired CTCF binding sites (13,675 sites, defined in Fig. 1 B), T_EFF-acquired concordant CTCF + ChrAcc sites (5,034 sites, defined in Fig. 1 C), and congruous CTCF sites (1,709 sites, defined in Fig. 6 C).