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. 2014 Nov 26;42(22):13949–13962. doi: 10.1093/nar/gku1247

Figure 3.

Figure 3.

mRNA transcripts encoding the BLV pri-miRNAs do not generate miRNAs. (A) Schematic diagram of the Rluc BLV 3′ UTR construct in which the BLV Gag-Pol 3′ UTR encompassing the miRNA cassette was cloned into Renilla luciferase expression vector. The terminator sequence at the 3′ end of each BLV pri-miRNA hairpin was mutated (X) to generate the Rluc BLV 3’UTR TM construct. Gray arrow indicates CMV promoter. (B) Semi-quantitative PCR amplification of the BLV miRNA-encoding region was performed from HEK293T cells transfected with either the Rluc BLV 3′ UTR or Rluc BLV 3′ UTR TM expression vectors to confirm expression the Rluc mRNA encoding the BLV pri-miRNAs. (C) Northern blot analysis from HEK293T cells transfected with either the Rluc BLV 3’ UTR or Rluc BLV 3’UTR TM expression vectors. Asterisks indicate putative RNAP III transcriptional read-through transcripts (*400 nt; **110 nt; ***100 nt; ****80 nt).