Fig. 4

Sub-cellular localization of BLV antisense transcripts. YR2 was fractionated into cytoplasmic and nuclear enriched pools of RNA, followed by production of total RNA and small RNA libraries for high throughput sequencing (HTS). Read count for each library was normalised and enrichment in the cytoplasm/nucleus calculated for the different classes of transcript. The HTS results were confirmed with RT-PCR using primers for the antisense BLV transcripts AS1 and AS2. Primers for TYR (cytoplasmic) and U6 (nuclear) were used to confirm fractionation efficacy. Both approaches confirm that the BLV antisense transcripts has a primarily nuclear localization (snoRNA small nucleolar RNA, snRNA small nuclear RNA)