MOESM1 of Recapitulation of gametic DNA methylation and its post-fertilization maintenance with reassembled DNA elements at the mouse Igf2/H19 locus
2020-01-15T05:12:52Z (GMT) by
Additional file 1: Figure S1. Generation and structural analysis of YAC-TgM carrying the 5′-truncated H19 ICR fragments. (A) Schematic representation of the YAC transgenes. The positions of the β-like globin genes (filled boxes) are shown relative to the locus control region (LCR, gray box). SfiI restriction enzyme sites are located 5′ to the LCR, within the LCR, and in the right arm of the YAC. Probes (filled rectangles) used for long-range structural analyses shown in panels (B), and the expected restriction enzyme fragments and their sizes are shown. The enlarged map shows the detailed structure of the del-8/9, 6/7, 4/5, and 2/3 fragments inserted between the LCR and the λ-globin gene. The positions of loxP5171 and loxP2272, inserted for employing the co-placement strategy, are indicated as solid and open triangles, respectively. (B) Long-range structural analysis of the transgenes in the YAC-TgM. DNA from thymus cells was digested with SfiI in agarose plugs and separated by pulsed-field gel electrophoresis, and Southern blots were hybridized separately to probes shown in (A). (C) In vivo Cre-loxP recombination in the parental del-8/9 transgene generates either del-8 or del-9 daughter transgenes. Positions of BamHI (B) restriction enzyme sites, and the expected restriction enzyme fragments and their sizes are shown. For example, if recombination occurs between the loxP5171 sites (solid triangles), no further recombination can occur because one of the loxP2272 sites (open triangles) is concomitantly deleted. The probe used for Southern blot analysis in (D) was shown as filled rectangles. The other TgM sub-lines were also generated by the same strategy. (D) Tail DNA from each YAC-TgM sublines was digested with BamHI and separated on agarose gels, and Southern blots were hybridized to the probe shown in (C).