Fig. 3

Pax7-engineered ESCs efficiently acquired the myogenic DNA methylation profile. a Experimental scheme of the differentiation protocol to generate ES-derived myogenic progenitors using iPax7 mES cells (left). Pax7 induction assessed by qRT-PCR (right). b Expression profiles of markers of pluripotency (Pou5f1 and Dppa4), muscle lineage commitment (Pax3, Myf5 and MyoD) and muscle differentiation (Myog, Ckm and Myh8) were measured by qRT-PCR at the successive time points of iPax7 ESC-derived myogenic model and normalized by Gapdh expression (n = 3, mean ± SD). c DNA methylation dynamics of Pax3, Myf5 and MyoD enhancers and CpG-poor promoters of muscle-specific genes (Myf5 Myog, Ckm and Myh8) and pluripotency genes (Pou5f1 and Dppa4) during Pax7-induced myogenesis (+Dox shown in red and -Dox shown in black). The results of three biological replicates are shown. Each circle represents a CpG dinucleotide and the colour gradient represents the level of methylation indicated in the legend and assessed by sodium bisulphite sequencing. Demethylation waves occurring during the Pax7-induced myogenesis are denoted by red dotted boxes. ESCs embryonic stem cells, Dox doxycycline; iPax7 inducible Pax7