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. 2009 Sep 15;37(20):6950–6959. doi: 10.1093/nar/gkp764

Figure 3.

Figure 3.

VA RNAI (A), but not VA RNAI (G), generates active RISC containing the 5′-strand of mivaRNAI. (A) Schematic representation of the terminal sequence of VA RNAI and the oligonucleotide used for primer extension. (B) Primer extension on total RNAs prepared from control 293-Ago2 cells (lane 1) or cells transfected with plasmid pVA RNAI(A) (lane 2) or pVA RNAI(G) (lane 3) or infected with virus Ad5 (lane 4) or wt900 infected cells (lane 5), were analyzed by primer extension for the transcription initiation start sites of VA RNAI. The products generated from ‘A’ start and ‘G’ start VA RNAI are indicated in the Figure. (C) The 293-Ago2 cells were transfected with plasmids pVA RNAI(A), pVA RNAI(G) or the empty vector as a control. The immunopurified RISC complexes from S15 cytoplasmic extracts were assayed for activity against the reverse VAI 5′ or VAI 3′ target RNAs.