(A to G) Day 20 post-differentiation neurons derived from WT and ADAR1 KO hESCs were transduced with lentivirus containing the indicated doxycycline-inducible shRNA. (A) 3′UTR shortening of selected genes measured by qPCR. The long 3′UTR transcript isoform was normalized to total ORF-containing transcripts. (B) Immunofluorescent images of dsRNA (J2, green) and DAPI (blue) in transduced WT neurons. Expression of doxycycline-inducible shRNA is coupled with red fluorescent protein (RFP) expression (red). (C) Quantification of dsRNA in (B). (D) Relative IFN-β mRNA levels measured by qPCR. (E to G)Same as (B) to (D) above, except in ADAR1 KO neurons. For all qPCR, RPS11 was used as a housekeeping gene. All quantified data shown are mean ±SD [n = 3 biological replicates in (A), (D), and (G); n = 12 cells from biological triplicates in (C) and (F)]. Two-way ANOVA with Tukey corrected multiple comparisons, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.