Scale bar 5 m b dna immuno fish of eraf red with rnap

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Scale bar, 5 m. ( b ) DNA immuno-FISH of Eraf (red) with RNAP II staining (green). ( c ) Comparison of the percentage of alleles exhibiting a gene transcription signal by RNA FISH (black), with the percentage of loci that overlap with an RNAP II focus by DNA FISH (gray) for Hbb-b1 ( n = 83), Eraf ( n = 59), Uros ( n = 47) and P2ry6 ( n = 79). Transcribing alleles localize to Polymerase II-rich structures called Transcription Factories Pol II Transcribing (RNA FISH) Eraf alleles 90% of alleles are active 90% of alleles are at trx factory Eraf gene (DNA FISH) alleles
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Double-label RNA FISH for Hbb-b1 gene transcripts and various other genes. Red signals are Hbb-b1 and green signals are Eraf ( a ), Uros ( b ), Igf2 ( c ), Kcnq1ot1 ( d ) and Hba ( e ). The percentage of signals that overlap with an Hbb-b1 signal is given at the bottom of each panel. DAPI staining is blue. Scale bar, 5 m. Transcription Factories some genes share sites of transcription, a possible gene “anchoring” site From Osborne et al. (2004) Nature Genetics 36, 1065-1071.
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Hbb Eraf RNA Pol II Transcribed genes can share a transcription factory
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Co-Association Frequencies of Transcribing Myc Alleles with Immunoglobulin Genes (A C) Double-label RNA FISH in stimulated B cells (10 min), showing Myc transcription signals in green, and in red, Igh (A), Igk (B), and Igl (C). DAPI staining is blue. Scale bar, 2 μ m. (D) The percentage of Myc signals that overlap with Igh, Igk, or Igl signals. One hundred nuclei were examined in each case. Myc - Igh versus Myc - Igk, p = 0.02; versus Myc - Igl, p = 0.0003. Frequency of Myc/Ig loci association mirror translocation frequencies in plasmacytomas these genes not associated in kidney or erythroid cells Osborne et al. (2007) PLoS Biol 5:e192 Myc - Chr 8 Igh - Chr 14
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from Sutherland & Bickmore (2009) Nature Rev Genetics 10, 457 Transcription in a Transcription Factory RNA PolII is stationary ??!!
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Schematic representation of chromatin loops (black) extruding from a chromosome territory (gray). Transcribed genes (white) in RNAP II factories (black circles). Potentiated genes (free loops) that are not associated with RNAP II factories are temporarily not transcribed. Potentiated genes can migrate to a limited number of preassembled RNAP II factories to be transcribed (dotted arrows). We propose that both cis and trans associations are possible.
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Attachments that may contribute to chromosome positioning may have functional consequences Parada and Misteli 2002, p. 425 Attachment and functional roles for chromosome positioning. (a) Tethering of chromosome territories to nuclear structures restricts their movement and contributes to maintenance of chromosome positioning. (b) Chromosome territories might be positioned within the nucleus in order to place specific genes in particular nuclear neighbourhoods that favour the expression or silencing of genes.
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Clustering of active genes at “transcription factories”; role of nuclear bodies? from Kosak and Groudine (2004) Science 306, 644
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Steven T. Kosak et al. Genes Dev. 2004; 18: 1371-1384
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