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 ) Structure of butyryloxymethyl-modified IP 6 (Pro-IP 6 ). ( B ) IP 6 levels in HeLa cells and HeLa IPPK KO cells C1 and C4 treated or untreated with 10 µM Pro-IP 6 for 24 h were analyzed using 36% polyacrylamide gel electrophoresis and toluidine blue staining. ( C ) Western blot analysis of ADAR1, ADAR2, IPPK and β-actin in HeLa cells and HeLa IPPK KO C1 and C4 cells not treated or treated with 10 µM Pro-IP 6 for 24 h. ( D ) Editing analysis of BLCAP and EEF2K editing sites in RNA from HeLa WT cells and HeLa IPPK KO C1 and C4 cells (n=3) untreated or treated with different concentrations of Pro-IP 6 for 24 h. ( E ) Alu editing index and inverted Alu in 3’UTR editing index of HeLa cells (WT) and HeLa IPPK KO (C4) untreated or treated with10 µM Pro-IP 6 for 24 h using NGS (n=3). ( F ) Editing analysis (heatmap) of some specific ADARs editing sites of HeLa cells (WT) and HeLa IPPK KO (C4) untreated or treated with10 µM Pro-IP 6 for 24 h using NGS data. Each data point represents one experiment. Statistical significance was analyzed using one-way ANOVA with Dunnett’s (D) or Tukey’s (E) post hoc test, *P < 0.05, **P < 0.01, or ***P < 0.001 compared to the WT cells without treatment (D) or cells untreated with Pro-IP 6 (E).

Article Snippet: Pro-IP 6 was synthesized as previously described , or acquire from TCI (Cat. # P3062).

Techniques: Modification, Polyacrylamide Gel Electrophoresis, Staining, Western Blot

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 ) Schematic representation of the RNA editing–dependent dual-luciferase reporter construct. ( B and C) Editing analysis of HeLa cells and HeLa IPPK KO C4 (n=3) untreated or treated with different Pro-IP 6 concentrations using the RNA editing–dependent dual-luciferase reporter assay. ( D ) Metabolic pathway of IP 6 synthesis and degradation by IPMK, IPPK and multiple inositol polyphosphate phosphatase 1 (MINPP1). ( E ) Western blot analysis of MINPP and GAPDH in HEK293 cells and HEK293 MINPP KO cells. ( F ) IP 6 analysis and quantification (n=5) in HEK293 cells and HEK293 KO MINPP cells by 36% polyacrylamide gel electrophoresis and toluidine blue staining. ( G ) Western blot analysis and quantification (n=3) of ADAR1 (p150+p110) and β-actin in HEK293 cells and HEK293 MINPP KO cells. ( H ) Editing analysis of HEK293 cells and HEK293 MINPP KO cells (n=3) using the RNA editing–dependent dual-luciferase reporter assay. Each data point represents one experiment. Statistical significance was analyzed using Student’s t test (H, L, M and N) or one-way ANOVA with Dunnett’s (I) post hoc test compared to WT cells without treatment, *P < 0.05, **P < 0.01, or ***P < 0.001.

Article Snippet: Pro-IP 6 was synthesized as previously described , or acquire from TCI (Cat. # P3062).

Techniques: Luciferase, Construct, Reporter Assay, Western Blot, Polyacrylamide Gel Electrophoresis, Staining

Journal: Journal of Neuroinflammation

Article Title: Systemic cytokines and GlycA discriminate disease status and predict corticosteroid response in HTLV-1-associated neuroinflammation

doi: 10.1186/s12974-022-02658-w

Figure Lengend Snippet: Transcriptomic validation of cytokine signaling pathways and prednisolone response in an independent cohort of PLHTLV-1. Digital transcriptomics (nCounter) was used to quantify cytokine signaling pathways in HAM/TSP disease progression, and the effect of in vitro prednisolone treatment in PBMCs from four AS, four HAM/TSP patients (including the two only incident HAM/TSP in the HOST cohort), and four age-, gender- and ethnicity-matched healthy controls. A Ex vivo IFN signaling score was significantly higher in incident HAM/TSP as compared to healthy controls, AS, and HAM/TSP at 4-year follow-up (one sample t test). B Ex vivo IL-17 signaling score was significantly higher in incident HAM/TSP as compared to healthy controls and tends to decline in HAM/TSP at 4-year follow-up (one sample t test). C IFN signaling was homogeneously down-regulated in all clinical groups by prednisolone treatment in vitro (left panel, Wilcoxon test p < 0.0001). Down-regulation was confirmed by decreased expression of the IFN-γ-regulated MHC Class II antigen presentation pathway (right panel, Wilcoxon test p < 0.001). D Ex vivo transcriptomic IFN signaling score measured by nCounter is significantly correlated with mRNA levels of previously identified HAM/TSP biomarkers CASP5, FCGR1A, STAT1, and CXCL10 (all p < 0.05 with Bonferroni correction), but not to HTLV-1 mRNAs Hbz and Tax. HC: healthy controls (open circles); AS asymptomatics (black circles), iHAM incident HAM/TSP (orange circles), HAM/TSP at 4-year follow-up (red circles), Con untreated in vitro PBMCs, Pred prednisolone-treated PBMCs in vitro. Paired samples from iHAM patients at diagnosis and at follow-up are identified by dashed lines.

Article Snippet: An additional biomarker, the chemokine CXCL10 (also known as IP-10) was quantified with the Bio-Plex Pro Human Chemokine IP-10/CXCL10 kit (Bio-Rad), following the manufacturer’s instructions [ ].

Techniques: In Vitro, Ex Vivo, Expressing