Figure 4. KPAF3 is necessary and sufficient to stabilize pre‐edited mRNAs.

1. Pre‐edited mRNA decay in KPAF3 RNAi cells at 36 h (left panel) and 48 h (right panel) post‐induction. After RNAi induction, Actinomycin D and ethidium bromide were added to inhibit transcription. Total RNA was isolated from cells collected at indicated time points after ActD/EtBr bromide addition, separated on denaturing 5% PAGE, and hybridized with DNA probe for pre‐edited RPS12 mRNA. Quantitation was performed in reference to 5.8S rRNA. The graphs below Northern blotting panels represent changes in relative abundance, assuming the mRNA/5.8S rRNA ratio at the time of ActD/EtBr addition as 100%. Contrast was increased in the right panel to reflect RNA loss at 48 h of KPAF3 RNAi.
2. Fully edited RPS12 mRNA decay in KPAF3 RNAi cells. Same membrane as in (A) was hybridized with a probe specific for a fully edited RPS12 mRNA.