Pre-miR-886 is distinct from vtRNAs. (A) Sequence alignment among pre-miR-886 and vtRNAs. The identical sequence blocks are highlighted in gray. The conserved A and B boxes are indicated by asterisks. (B) Northern hybridization and Western blotting in the five cell lines used in Figure 1D (which contains the loading control for the Northern). (C) qRT-PCR measurement of vtRNAs in the five oral cell lines as described in Figure 1E. (D) Western blotting (top two panels) and Northern hybridization (middle three panels) in subcellular fractions P10 (the nuclear pellet), S100 (the cytoplasmic soluble fraction), and P100 (the cytoplasmic pellet). The same portion from each fraction was loaded and EtBr staining (bottom panel) was included for comparison between fractions. In Northern hybridization, total RNA (tot) was included. In samples in the right half of the panel, WI-38 lung fibroblasts were treated with doxorubicin (4 μg/mL) for 4 h before cell harvest.
