Characterization of dynamic CTCF binding sites in early T _EFF compared to T _N cells. (A) Gating strategy to identify CD45.2⁺ donor-derived P14 CD8⁺ T cells in WT CD45.1⁺ recipients. (B) PCA of CTCF binding profile in WT T_N and early T_EFF cells, where normalized CTCF CUT&RUN signals on merged peaks were used as input data. (C) Distribution of dynamic CTCF binding sites in promoter (TSS+/−1 kb) and distal regulatory regions, where values in bars denote actual site numbers. (D and E) Functional annotation of T_EFF-acquired (D) and T_EFF-lost (E) CTCF binding sites based on GREAT analysis using the TSS+/−100 kb association rule. Non-redundant GO terms were selected from top 20 Hyper Rank for display, where values in bars denote site numbers and orange bars denote specific connection with immune functions. (F) Top three motifs in T_N and T_EFF non-differential CTCF binding sites based on HOMER analysis, with the known motif results shown. (G and H) Top motifs in T_EFF-acquired (G) and T_EFF-lost (H) CTCF binding sites based on HOMER analysis, with the de novo motif results shown.