Figure 2. Knockdown of RPC6 induces a decrease in BLV miRNAs expression.

The 293T cell line was transduced with lentiviral vectors expressing different shRNAs directed against RPC6 or a non-targeting shRNA and then transiently transfected with a plasmid expressing the miRNA cluster (pmiR). Forty-eight hours post-transfection, total protein and total RNA were extracted. (a) The α-tubulin (upper panel, loading control) and RPC6 (lower panel) were immunodetected by western blotting. Relative quantifications are presented as percentages of RPC6 level in comparison with the non-targeting shRNA. (b–e) Quantitative real-time RT-PCR experiments were performed to measure BLV-miR-B4-3p (b), BLV-miR-B2-5p (c), cellular miR-191 (d) or Firefly luciferase expression (e). Values were normalized using GADPH gene primers. Data are the means ± SEM from one representative of at least two independent experiments.