Intrinsic Epigenetic Regulation of the D4Z4 Macrosatellite Repeat in a Transgenic Mouse Model for FSHD
Figure 1
Integration site and copy number of D4Z4-2.5 and D4Z4-12.5 constructs in the mouse genome.
A) Schematic draw of the L42 EcoRI fragment used to generate the D4Z4-2.5 mouse line B) Metaphase spread of D4Z4-2.5 fibroblasts co-stained with dapi and the CY3 labeled L42 probe shows integration at a single pair of chromosomes C) COBRA-FISH analysis on D4Z4-2.5 fibroblast metaphase spreads probed with biotinylated-L42 fragments shows integration of L42 on chr17; D) Detection of copy number of the integrated fragments in both mouse models by MLPA analysis. The probe mix contained three probes specific for wild type alleles, one probe designed against the human p13E-11 region and one probe against D4Z4 E) Schematic draw of PAC clones used to generate the D4Z4-12.5 mouse; F) COBRA-FISH analysis on D4Z4-12.5 fibroblast metaphase spreads probed with a biotinylated PAC clone shows integration of the PAC clone on chr2; G) Fiber-FISH analysis of D4Z4-12.5 fibroblasts. Both PAC clones, labeled and hybridized to DNA fibers, were shown to be recombined during integration into the mouse genome.