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Figure 2 | Epigenetics & Chromatin

Figure 2

From: Murine esBAF chromatin remodeling complex subunits BAF250a and Brg1 are necessary to maintain and reprogram pluripotency-specific replication timing of select replication domains

Figure 2

Role of BAF250a in the regulation of replication domains in cells undergoing somatic cell reprogramming. (A) Number of alkaline phosphatase (AP)-positive colonies derived from 4-hydroxytamoxifen (OHT)-treated BAF250a flox/flox and BAF250a flox/flox; Mer-Cre-Mer mouse embryonic fibroblasts (MEFs) after Oct4, Sox2, Klf4, and c-Myc (OSKM) expression. (B) Morphology of AP-positive colonies derived from BAF250a flox/flox and BAF250a flox/flox; Mer-Cre-Mer MEFs. (C) Replication timing profiles of control embryonic stem cells (ESCs) (mock-treated ESCs from Figure 1), BAF250a flox/flox OSKM, BAF250a -/- OSKM, and MEFs. Replication timing was analyzed in two independent BAF250a flox/flox OSKM clones and three independent BAF250a -/- OSKM clones, and the averaged data are shown for each genotype. (D) Correlation analysis between replication timing datasets (Pearson’s R values). (E) Replication timing changes of BAF250a-affected domains in OSKM cells (BAF250a -/- ratio - BAF250a flox/flox ratio) were plotted against the same changes in ESCs (OHT ratio - mock ratio). (F) Summary of significant early to late (EtoL) and late to early (LtoE) switching segments in OHT-treated cells determined as in Figure 1C. (G) Venn diagram showing the overlap between genomic segments that undergoes EtoL switching (top) and LtoE switching (bottom) upon BAF250a loss in ESCs versus OSKM cells (FDR = 1% segments from Figure 1C and Figure 2F). (H) Replication timing plots of exemplary EtoL and LtoE switching domains.

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Epigenetics & Chromatin

ISSN: 1756-8935