rd systems elisa kit  (R&D Systems)

Journal: Molecular Therapy Oncology

Article Title: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Elevated transaminases and toxicity from high plasma mIFNβ levels after hydrodynamic injection of plasmid DNA encoding mIFNβ in Balb/c mice (A) Schematic representation of the sleeping beauty transposon system with a transposon plasmid pKT2/mIFNβ-CLP PGK-Luc encoding mIFNβ, and sleeping beauty SB100X transposase. (B) Plasma IFN-β levels from Balb/c mice [0.0037 μg DNA ( n = 5), 0.011 μg DNA ( n = 3), 0.033 μg DNA ( n = 3), 0.05 μg DNA ( n = 3), 0.1 μg DNA ( n = 4), 0.2 μg DNA ( n = 3), 0.4 μg DNA ( n = 6), 0.8 μg DNA ( n = 3), and 1.6 μg DNA ( n = 1)] at 6–10 days after hydrodynamic injection of increasing amounts of plasmid DNA. Mean ± SEM. (C) AST and (D) ALT levels of individual mice with the corresponding plasma mIFNβ are plotted. The respective Pearson correlation factor and p values are shown. High systemic IFN-β is associated with elevated AST and ALT.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Clinical Proteomics, Injection, Plasmid Preparation

Journal: Molecular Therapy Oncology

Article Title: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Lack of toxicity in the absence of IFNR signaling and toxicity is reversible after IFN-β production is terminated Transgenic mice knockout for type I IFN receptor (IFNRko) and C57/B6 mice received one intramuscular injection of saline or 2 × 10 11 vg AAV1-mIFNβ and were monitored over time. (A) Plasma mIFNβ, (B) Kaplan-Meier survival curves, (C) ALT, (D) AST from saline (black, n = 6), or 2 × 10 11 vg AAV1-mIFNβ-treated IFNRko mice (red, n = 10), and C57B/6 mice (blue, n = 7) were plotted at regular intervals. Line denotes mean. Half of the mice were bled for each time point for blood analysis. C57/B6 mice ( n = 25) were given one intramuscular injection of AAV-mIFNβ-iCas9 and 14 days later, given one intraperitoneal dose of AP1903 a prodrug to activate iCas9 to initiate apoptosis of AAV-transduced muscle cells expressing IFN-β. (E) Plasma mIFNβ, (F) ALT, (G) AST, and (H) platelet counts in C57B/6 mice before (day 0) and after termination of mIFNβ production using AP1903. Mean ± SEM.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Transgenic Assay, Knock-Out, Injection, Saline, Clinical Proteomics, Expressing

Journal: Molecular Therapy Oncology

Article Title: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Concomitant treatment with ruxolitinib protects VSV-IFNβ-NIS treated MPC-11 tumor-bearing Balb/c mice from toxicity (A) Kaplan-Meier survival, (B) individual tumor volumes, (C) predicted tumor volumes using mixed effects modeling, (D) plasma IFNβ, (E) ALT, (F) AST, and (G) platelet count of mice at scheduled bleeds on day 3 and day 14, or at necropsy (at various time points). Treatment groups are color coded, saline (black, n = 5), VSV alone (red, n = 10), ruxolitinib (gray, n = 5), and VSV plus ruxolitinib (blue, n = 10). One intravenous dose of VSV at 10 7 TCID 50 was given per mouse, and/or ruxolitinib at 2 mg twice a day by oral gavage for 10 days, starting on the same day as VSV therapy.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Clinical Proteomics, Saline

Journal: Advanced Science

Article Title: RNA‐Binding Protein RBM25 Targets the mRNA Stability of GTPase Rab22a to Restrict Viral Entry and Infection

doi: 10.1002/advs.76160

Figure Lengend Snippet: RBM25 mediates antiviral protection independent of the IFN‐I signaling pathway. (A) A scheme of RNA‐seq analysis of WT and Rbm25 ‐cKO peritoneal macrophages infected with VSV for 4 h. (B) Heatmap of differentially expressed genes (DEGs) in Rbm25 ‐deficient and WT peritoneal macrophages infected with VSV for 4 h. (C) GSEA showing enrichment of DEGs in the pathways involving in the response to IFN‐α, IFN‐β production, JAK‐STAT3, IFN‐I receptor binding, toll like receptor, and cGAS target gene signaling pathways. (D) RT‐qPCR analysis of IFN‐I ( Ifna4, Ifnb1 ) mRNA in the lung tissues from WT and Rbm25 ‐cKO mice infected with PR8 influenza virus with a sublethal dose (50 PFU., n = 5 per group) at 7 dpi. (E) RT‐qPCR analysis of Cxcl10, Mx1, Isg15 , and Mx2 mRNA in the lung tissues from WT and Rbm25 ‐cKO mice treated as in (D). (F) ELISA of IFN‐β in serum from Rbm25 ‐cKO and WT mice infected with PR8 influenza virus, as in (D). (G) RT‐qPCR analysis of IFN‐I ( Ifna4, Ifnb1 ) mRNA in peritoneal macrophages from Rbm25 ‐cKO and WT mice infected with PR8 influenza virus for 12 h. (H) RT‐qPCR analysis of Cxcl10, Isg15 , and Rsad2 mRNA in peritoneal macrophages from WT and Rbm25 ‐cKO mice treated as in (G). (I) Immunoblots analysis of RBM25, RIG‐I, MAVS, p‐TBK1, TBK1, p‐IRF3, and IRF3 in peritoneal macrophages from WT and Rbm25 ‐cKO mice infected with PR8 influenza virus for the indicated times. β‐ACTIN was used as a loading control. (J) A scheme of experiment analyzing the infection of viruses in peritoneal macrophages from Irf3 −/− mice transfected with control siRNA or siRNA targeting Rbm25 . (K) RT‐qPCR analysis of PR8 HA and IAV M1, HSV‐1 TK and ICP0, VSV‐G mRNA in peritoneal macrophages from Irf3 −/− mice transfected with control siRNA or siRNA targeting Rbm25 followed by infection with PR8 influenza virus for 12 h, HSV‐1 for 12 h, or VSV for 8 h, respectively. (L, M) RT‐qPCR analysis of IFN‐I ( Ifna4, Ifnb1 ) mRNA in Rbm25 ‐cKO and WT peritoneal macrophages transfected with poly (I:C) for 4 h (L) or stimulated with UV‐VSV for 18 h (M). (N) Immunoblots analysis of p‐STAT1 and STAT1 in peritoneal macrophages from WT and Rbm25 ‐cKO mice infected with PR8 influenza virus for the indicated times. β‐ACTIN was used as a loading control. Data are presented as the mean ± SD. Unpaired two‐tailed Student's t ‐test (D–H, K–M). ns, not significant.

Article Snippet: IFN‐β levels in the supernatants or sera were measured using a mouse IFN‐β ELISA kit (PBL Biomedical Laboratories) according to the manufacturer's instructions.

Techniques: RNA Sequencing, Infection, Binding Assay, Protein-Protein interactions, Quantitative RT-PCR, Virus, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Transfection, Two Tailed Test