Isolation of endogenous ADAR-associated RNAs. (A) Agarose gel of RNA isolated from total egg extract, mock immunoprecipitation or ADAR immunoprecipitation. (B) Histogram of transcript density as a function of ADAR enrichment. (C) RT-PCR for predicted ADAR-enriched transcripts from total egg extract or an ADAR immunoprecipitation. PP4R1, a transcript predicted not to be ADAR-enriched, serves as a negative control. Results shown are from one representative experiment. Similar results were obtained from a total of three biological replicates. (D) Charts representing the fraction of total transcripts, transcripts predicted to form >15 bp hairpins, and transcripts predicted to form >200-bp intermolecular dsRNA that have ADAR-enrichment values ≥4. P-value for overrepresentation of ADAR-enriched transcripts among transcripts predicted to form hairpins is 1 × 10−14 using a one-sided Fisher's exact test. P-value for overrepresentation of ADAR-enriched transcripts among transcripts predicted to form intermolecular dsRNA is <2.2 × 10−16 using a one-sided Fisher's exact test.
