Journal: iScience

Article Title: Identification of ADAR1i-124: The first effective A-to-I RNA editing inhibitor with promising cancer therapeutic potential

doi: 10.1016/j.isci.2025.114615

Figure Lengend Snippet: Detection of unedited and/or under-edited dsRNAs and Z-RNAs induced by ADAR1i-124 (A) Immunostaining with the dsRNA-specific mAb J2 revealed unedited and/or under-edited dsRNAs in Yumm1.7 cells, exclusively induced by compounds (10 μM) that inhibit RNA editing. Scale bars, 10 μm. (B) The unedited and/or under-edited dsRNAs induced by siAdar1 and ADAR1i-124 were sensitive to RNase III treatment. Scale bars, 50 μm. (C) Co-localization of unedited and/or under-edited dsRNAs and MDA5 detected by immunostaining using both J2 and anti-MDA5 antibodies. Scale bars, 20 μm. (D) Immunostaining with the Z-RNA-specific mAb Z22 showed that certain dsRNAs underwent a Z-form transition in Yumm1.7 cells treated with siAdar1 and ADAR1i-124. The Z-RNAs induced were also susceptible to RNase III degradation. Scale bars, 50 μm. (E) RNA-seq analysis of Yumm1.7 cells treated with ADAR1i-124 (10 μM 12 h) versus 0.1% DMSO (control) was conducted to assess global A-to-I editing changes. Sites mapped with >20 reads in both samples and >1 edited reads in either sample are plotted. Sites exhibiting significant differences in editing levels ( p < 0.05, χ 2 test) between the ADAR1i-124 treatment and DMSO-control are colored in green for regular editing sites and in orange for hyper-edited sites. Sites exhibiting insignificant differences are colored in gray. (F) Types of transposable elements exhibiting significantly lower RNA editing levels upon ADAR1i-124 treatment. Significantly inhibited sites by ADAR1i-124 ( n = 910) are annotated by RepeatMasker for TE elements.

Article Snippet: IB: anti-MDA5 , Cell Signaling Technology , 5321.

Techniques: Immunostaining, RNA Sequencing, Control