Fig. 8: Identification of CTCF & Suz12 binding in 19 kb silencer element and consequences of CTCF binding site deletion.
From: An oligodendrocyte silencer element underlies the pathogenic impact of lamin B1 structural variants
UCSC genome browser tracks of CUT&RUN analysis of CTCF in (a) human fibroblast, (b) mouse ES cells, (c) mouse primary oligodendrocytes (OL), and (d) SUZ12 in mouse primary OLs confirm bioinformatically predicted CTCF binding sites within 19 kb silencer element, and that SUZ12 associates with the CTCF2 region. Arrows point to the conserved CTCF 1 and CTCF 2 sites. Black bars above traces represent called peaks. e Schematic showing deletions of CTCF 1 and 2 sites. Real-time PCR analysis of Lmnb1 mRNA expression in Oli-neu, N2A and 3T3 cells with CRISPR-mediated deletions of (f) CTCF1 and (g) CTCF2. Lmnb1 expression is significantly higher in Oli-neu cells with CTCF1 deleted but lowered in N2A and 3T3 cells, relative to their respective controls. Lmnb1 expression is increased in Oli-neu and N2A cells with CTCF2 deleted but lowered in 3T3 cells. Lmnb1 is normalized to β Actin (Actb). Graphs are mean ± SEM. For CTCF1 - Oli-neu, control & del n = 3; N2A, control n = 5, del n = 3; 3T3, control n = 4, del n = 5. For CTCF2 - Oli-neu, control n = 6, del n = 4; N2A, control n = 6, del n = 4; 3T3, control n = 5, del n = 4. In all cases, independent clones were used. *p < 0.05, **p < 0.01, two-tailed tailed t-tests.
