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. 2009 Nov 26;5(11):e1000739. doi: 10.1371/journal.pgen.1000739

Figure 8. Biallelic IGF2 expression after cohesin depletion.

Figure 8

(A) Sequencing of genomic DNA (gDNA) shows that the HB2 cell line is heterozygous for a SNP in IGF2. Pyrosequencing of cDNA of cohesin depleted G2 phase cells shows both IGF2 alleles indicating biallelic IGF2 expression. (B) Sequencing of genomic DNA (gDNA) shows that the HB2 cell line is heterozygous for a SNP in H19. Pyrosequencing of cDNA of cohesin depleted and control G2 cells shows only the C allele indicating that H19 expression remains monoallelic after cohesin depletion. (C) After labelling IGF2 RNA by RNA fluorescence in situ hybridisation (FISH; green) on cohesin depleted (SCC1 RNAi) and control interphase HB2 cells (RNAi control), DNA FISH (red) was performed to detect genomic IGF2 sequence. Nuclei were counterstained with DAPI (blue) and imaged by confocal microscopy. Representative images are shown; first panel: overlay, second panel: DNA signal, third panel: RNA signal. IGF2 expression increases and becomes biallelic upon cohesin depletion.