Integration of a transgenic <i>β^A</i> replicator flanked by insulators induces early replication timing at two other chromosomal loci.

<p>Replication timing profiles following transgene integration into erythroid 6C2 cells. (A) Schematic representation of the <i>IL-2R</i> transgene drawn to scale. The construct is identical to the one described in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001277#pbio-1001277-g005" target="_blank">Figure 5A</a>, except that HS4 carries deletions of the CTCF (ΔFII) or VEZF1 (ΔFIII) binding sites. This construct is linked to the hygromicin antibiotic resistance gene under the control of the HSV-thymidine kinase promoter. (B) The mapped sites of integration (red dots) for the transgenes flanked by ΔFII or ΔFIII insulators are shown. UCSC genome browser views (May 2006 build) of 100 kb regions with the chromosomal position, GC percentage, annotated genes, and CpG islands shown. (C) Replication timing profiles of chromosomal alleles at the mapped sites of transgene integration. Late and early replicated controls correspond to the endogenous <i>amylase</i> α 2A and <i>β-globin</i> loci, respectively. The differences in late or early replication (ΔL and ΔE) at flanking regions compared with the integration site are shown.