JCS02945 Supplementary Material

Files in this Data Supplement:

• Supplemental Tables - Adobe PDF

• Supplemental Figure 1 -

  Figure S1. Early redistribution of heterochromatin in hB3C2 heterokaryons. CREST immunostaining was used to compare the distribution of constitutive heterochromatin in human B lymphocyte (hB) derived nuclei before fusion (d0), 2 days (d2) and 7 (d7) days after hB3C2 heterokaryon formation and successful reprogramming (5.1H11-positive). The number of chromocentres and the number of discrete CREST signals was assessed using 3D-reconstructed sequential z-series spanning 100 individual nuclei for each time point. Chromocentres were defined as three or more closely juxtaposed CREST signals and differentiating C2C12 cultures (0, 2 and 7 days) were analysed as controls (right panel).

• Supplemental Figure 2 -

  Figure S2. hPAX5 extinction in heterokaryons is not associated with a decline in the expression of hE2A and hEBF. qRT-PCR analysis of human hPAX5, hE2A and hEBF expression in hBxC2 heterokaryons treated with 20 nM TSA (white bars) or untreated (black bars). Expression of hE2A and hEBF was compared with control cultures of differentiating human HFM (Human foetal muscle) cells (grey bars). Mouse C2C12 samples (C2) were used as negative controls to confirm the specificity of primers for human transcripts and the data were normalised to hGAPDH expression. The sequence of human specific pairs of primers used for PCR amplification is available upon request.