Figure 2.

Alteration of single/few nucleotide(s) in M1 or M2 dramatically impact(s) the binding of CTCF. (A) The wild-type (WT) and 3 bp mutant (Mut) sequences of CTCF binding M1. (B) Comparisons of the binding capacity of WT M1 and mutant M1. The binding of GST-CTCF-ZFs fusion protein to WT M1 (lane 1), to Mut 1 (from TGG to GTT) (lane 2), to Mut 2 (from TGC to TAA) (lane 3), to Mut 3 (from CCT to AAA) (lane 4), to Mut 4 (from TGC to GTA) (lane 5), to Mut 5 (from TGG to GTT) (lane 6), to Mut 6 (from CCA to AAC) (lane 7), and to Mut 7 (from TCT to AAG) (lane 8). (C) The WT and 2 bp Mut sequences of CTCF binding M1. (D) Comparisons of the binding capacity of WT M1 and 2 bp Mut M1. The binding of GST-CTCF-ZFs fusion protein to WT M1 (lane 1), to Mut 8 (from TG to GT) (lane 2), to Mut 9 (from GG to TT) (lane 3), and to Mut 10 (from GT to TG) (lane 4). (E) The WT and 1 bp Mut sequences of CTCF binding M1. (F) Comparisons of the binding capacity of WT M1 and 1 bp Mut M1. The binding of GST-CTCF-ZFs fusion protein to WT M1 (lane 1), to Mut 11 (from T to G) (lane 2), to Mut 12 (from G to T) (lane 3), and to Mut 13 (from G to A) (lane 4). (G) The WT and 1 bp Mut sequences of CTCF binding M2. (H) Comparisons of the binding capacity of WT M2 and 1 bp Mut M2. The binding of GST-CTCF-ZFs fusion protein to WT M2 (lane 1), to Mut 1 (from A to T) (lane 2), to Mut 2 (from T to G) (lane 3), and to Mut 3 (from C to A) (lane 4). (I) Comparisons of the binding capacity of CTCF to both WT M1 and Mut M1 by ChIP-qPCR. (J) Comparisons of the binding capacity of CTCF to both WT M2 and Mut M2 by ChIP-qPCR