Journal: bioRxiv

Article Title: Inositol hexakisphosphate Functions as a Cofactor and Modulator of ADAR1 Activity

doi: 10.64898/2026.01.17.699700

Figure Lengend Snippet: ( A ) Scheme of ADAR1 protein, ADAR1 in vitro translated construct (ADAR1-IVTra) and the experimental setup for protein generation. ( B ) In vitro synthesized ADAR1-IVtra using no IPs or IP 6 or IP 5 (2-OH-IP 5 ), were analyzed by SDS-PAGE and Coomassie staining before and after purification with MagStrep “type3” XT beads. Dihydrofolate reductase (DHFR) was used as control of the in vitro translation synthesis. ( C ) Schematic representation of the in vitro editing reaction using ADAR1-IVTra and different RNAs ( D-F ) In vitro editing reaction (n=3) of in vitro transcribed GLI1 , HTR2C and NEIL1 dsRNA using ADAR1-IVTra; secondary structure of the dsRNAs fragments (editing sites are depicted by arrows and A, B and C) were predicted using RNAfold; editing levels were analyzed by cDNA synthesis follow up by PCR amplification and sanger sequencing. ( G ) ADAR1-IVTra proteins used in the in vitro reactions of ( D - F ) were analyzed by SDS-PAGE and Coomassie staining. ( H ) Quantification of ADAR1-IVTra protein using different concentrations of IP 6 and IP 5 (left graphic), and corresponding editing of GLI1 B editing site after 1 h of in vitro editing reaction (right graphic). Each data point represents one experiment or one donor. Statistical significance was analyzed using one-way ANOVA with Dunnett’s post hoc test, *P < 0.05, **P < 0.01, or ***P < 0.001 compared to ADAR1-IVTra-no IPs.

Article Snippet: ADAR proteins were purified from the reaction mixture using MagStrep “type3” XT beads (Cat. # 2-4090-002, IBA) following the manufacturer’s instructions.

Techniques: In Vitro, Construct, Synthesized, SDS Page, Staining, Purification, Control, cDNA Synthesis, Amplification, Sequencing