13072_2016_56_MOESM5_ESM.tif (692.31 kB)
MOESM5 of Chicken embryonic stem cells and primordial germ cells display different heterochromatic histone marks than their mammalian counterparts
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posted on 2016-02-10, 05:00 authored by Clémence Kress, Guillaume Montillet, Christian Jean, Aurélie Fuet, Bertrand PainAdditional file 5. Western and dot blot supplementary data. (A) Western blot images of H3 post-translational modifications detection in chicken ESCs, RA-differentiated ESCs, BCs, PGCs and CEFs. 3 μg of purified histones were analysed by SDS-PAGE followed by blotting and immunodetection using an antibody against a modification (upper panels), and an antibody against H3 after stripping of the membrane (lower panels). Molecular weights are indicated in kDa. (C) Global levels of 5-methylcytosine and 5-hydroxymethylcytosine in mouse cells. 5mC and 5hmC levels were quantified by dot blot analysis of 125 ng of denatured genomic DNA from mouse ESC cultured in 2i or serum medium and from mouse epiblast stem cells (EpiSCs); pBSK plasmid was used as a negative control. Error bars indicate the standard deviation of the mean signal for three technical replicates of one representative experiment.