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Fig. 6 | Retrovirology

Fig. 6

From: Characterization of novel Bovine Leukemia Virus (BLV) antisense transcripts by deep sequencing reveals constitutive expression in tumors and transcriptional interaction with viral microRNAs

Fig. 6

Mutant proviruses altered in the microRNA region reveal interactions between the antisense transcripts and the viral microRNAs. a Ideograms showing the three BLV proviruses used in transfections, the first a wild-type, the second where the microRNAs were deleted (grey box indicates the region removed), the third where the BLV microRNAs were inverted. 3′RACE on total RNA from HeLa cells transfected with the three constructs shows that the microRNA deletion/inversion increases the fraction of AS1-L reads. In the case of the wild-type provirus and the microRNA-deleted provirus, reads were mapped to the wild type provirus sequence. In the case of the microRNA-inverted provirus, they were mapped to a proviral genome with microRNAs inverted to match the provirus used. b Nuclear versus cytoplasmic enrichment for BLV sense and antisense transcripts obtained via real-time RT-PCR with nuclear and cytoplasmic enriched RNA from HeLa cells transfected with either the wild type BLV pBLV344 provirus, the provirus with the BLV microRNAs deleted or with the orientation of the BLV microRNAs inverted. Removing or inverting the BLV microRNAs had no effect on the localization of the sense or antisense transcripts. TYR cytoplasmic enriched control, U6 nuclear enriched control

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