Journal: PLoS Genetics
Article Title: Induction of a chromatin boundary in vivo upon insertion of a TAD border
Figure Lengend Snippet: Recruitment of architectural proteins and topological changes upon integration of the TgN(38–40) construct. (A) ChIP of CTCF and RAD21 in wild-type or TgN(38–40) E12.5 whole limbs. The window displayed corresponds to the native region CS38-40. Dashed lines are displayed for better comparison between the occupancy of various CTCF sites. Peak calling is represented as black boxes. Bottom, extension of both the TgN(38–40) construct and del(CS38-40) background. (B) Hi-C showing the host locus of chromosome 10 in wild-type whole limbs at E12.5. Below the Hi-C heatmap, wild-type CTCFs (red or blue arrowheads) and topological domains (horizontal bars). (C) Distribution of H3K27ac (green) over the host locus in the distal part of wild-type E12.5 forelimbs. (D) Top, 4C-seq using CS38 and CS40 as viewpoints in TgN(38–40) E12.5 limbs. Bottom, 4C-seq tracks of CTCF-left and CTCF-right viewpoints in both TgN(38–40) homozygous (red lines) and wild-type samples (blue lines). Percentages of 4C-seq contact changes beyond the integration site are shown. Black arrowheads indicate 4C-seq viewpoints. (E) ChIP of CTCF and RAD21 over the host landscape in TgN(38–40) .
Article Snippet: Antibodies (CTCF, Active Motif 61311 or RAD21, Abcam ab992) were incubated with Dynabeads Protein A (Thermo Fisher Scientific, 10001D) for 3 hours on a rotating wheel at 4°C.
Techniques: Construct, Chromatin Immunoprecipitation, Hi-C