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Fig. 5. Coexpression of lymphocyte- and muscle-specific genes in individual nuclei treated with HDAC inhibitors TSA and VPA. (A) Human B lymphocytes were analysed for hCD20 (green) and hNCAM (red) transcripts using RNA FISH. Two foci of hCD20 signal are evident in hB cell nuclei that were removed by pre-treatment of samples with RNAse A (+RNase), but not DNAse (+DNAse). Application of 20 nM TSA did not impair hCD20 signal detection (+TSA). (B) Human NCAM transcripts (red) were detected in human myoblast nuclei but not in lymphocytes. (C) RNA FISH showing examples of nuclei that express hCD20 transcripts alone (green) and hNCAM transcripts alone (red) or both in lymphocyte-derived nuclei within day 5 hBxC2 heterokaryons. Confocal images are maximal projection of multiple optical z-sections (Bars, 10 µm). (D) the proportion of nuclei expressing hNCAM (red), hCD20 (green), or both hNCAM and hCD20 (red/green) signals in heterokaryons cultured in the absence (untreated) or presence of 20 nM TSA or 1 mM VPA is expressed as a proportion of human nuclei in which RNA transcripts were detected (see supplementary material Table S1).
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