TbRGG2 differentially affects editing of pan-edited A6 RNA at the 3′ and 5′ ends. (A) Diagram of fully edited A6 RNA depicting the nucleotide position (top) and the number of edited sites (ESs; bottom) on the fully edited RNA. The regions of the RNA where editing is examined using qRT-PCR and poisoned primer extensions (PPEs) are labeled. (B,C) Representative initiation PPEs (B) and 3′ end PPEs (C) were performed as described in Figure 2 but with primers specific to A6. (D) Comparison of the decrease in edited RNA levels following TbRGG2 loss using qRT-PCR to analyze 5′ fully edited transcripts (Fisk et al. 2008) and PPEs to analyze editing at initiation and near the 3′ ends of the transcripts. For the PPE reactions, the decreases in edited RNA levels were determined using RNA isolated from at least duplicate experiments. Edited RNA levels were normalized to tubulin PPEs performed in the same reaction.
