Figure 3

Nucleo-cytoplasmic distribution of Apobec-1-dependent mRNA editing targets. (A,B) Distribution of WT small intestine (A) and hepatic (B) edited apoB RNA. A 738 bp amplicon (nucleotides 6,508 to 7,246) from nuclear and cytoplasmic apoB mRNA was cloned and sequenced. Twenty-two clones from each subcellular fraction (from three independent nuclear-cytoplasmic isolations) were analyzed. Left panel: graphic representation of percentage of edited clones in nuclear and cytoplasmic apoB RNA. Right panel: targeted cytidines identified in nuclear apoB RNA are indicated with green circles; cytidines identified in cytoplasmic apoB RNA are represented by blue circles. All cytidines are aligned with the nucleotide position to the left. (C) Nuclear-cytoplasmic distribution of intestinal Apobec-1 3′ UTR targets identified by RNA-seq and validated by Sanger sequencing. A 550 bp (ATP6ap2) and a 667 bp (Usp25) amplicon were generated from nuclear and cytoplasmic RNA and analyzed by sequencing 19 to 22 clones. For both ATP6Ap2 and Usp25 RNAs, the edited RNA is predominantly exported to the cytoplasm.