Distribution of 5-hydroxymethylcytosine (5 hmC) and 5-methylcytosine (5 mC) in Ad-MSCs.

<p>Basal mRNA levels of (A) <i>TET2</i> and (B) <i>TET3</i> were determined by qRT-PCR in Ad-MSCs from young and old donors (each group: N = 6, n = 3). (C) Expression changes of <i>TET2</i> and <i>TET3</i> were confirmed on the protein level by Western blot analysis (N = 3/group). For both techniques <i>GAPDH</i> was used for normalization. (D) Young and old Ad-MSCs (each group: N = 3, n = 5) were stained for 5-hydroxymethylcytosine (5 hmC/red) and 5-methylcytosine (5 mC/green). Cell nuclei were counterstained with Hoechst 33342 (blue). (E&F) For quantification 5 hmC or 5 mC positive nuclei were counted. *p<0.05, **p<0.01 (student’s t-test).