mouse interferon beta ifn β enzyme linked immunosorbent assay  (Elabscience Biotechnology)

Journal: Cell reports

Article Title: Bilirubin metabolism in the liver orchestrates antiviral innate immunity in the body.

doi: 10.1016/j.celrep.2025.115481

Figure Lengend Snippet: Figure 2. UGT1A1 promotes IFN-I production and exerts broad-spectrum antiviral effects (A) RT-qPCR analysis of SeV RNA in HepG2 cells transfected with control shRNAs (shCtrl) or UGT1A1 knockdown plasmid shUGT1A1 (1# or 2#) and then infected with SeV (MOI, 1.0) for 24 h. (B) RT-qPCR analysis of UL42 RNA levels in Huh7 cells transfected with control shRNAs (Ctrl) or UGT1A1 knockdown plasmid shUGT1A1 (1# or 2#) and then infected with HSV-1 (MOI, 1.0) for 12 h. (C and D) Western blot detection of VSV-encoded G protein (VSV-G) (C) or H1N1-encoded HA protein (D) in HepG2 cells transfected with shCtrl or shUGT1A1 (1# or 2#) and then infected with VSV (MOI, 1.0) or H1N1 (MOI, 1.0) for 24 h. (E) HepG2 cells were transfected with control vectors or FLAG-UGT1A1 and then infected with VSV (MOI, 1.0) or H1N1 (MOI, 1.0) for 24 h. RT-qPCR was used to detect virus RNA. (F) Fluorescence microscopy of the VSV virus with a GFP gene (VSV-GFP) in HEK293T cells transfected with FLAG-UGT1A1 and then infected with VSV-GFP (MOI, 1.0) for 24 h. Scale bars, 100 mm. (G and H) RT-qPCR analysis of Ifnb mRNA in HepG2 cells transfected with FLAG-UGT1A1 and then infected with SeV (MOI, 1.0) for 12 h (G) or in Ugt1a1+/+ or Ugt1a1/ HepG2 cells infected with VSV (MOI, 1.0) for 12 h (H). (I) ELISA analysis of IFNb protein levels in Ugt1a1+/+ or Ugt1a1/ HepG2 cells infected with VSV (MOI, 1.0) for 12 h.

Article Snippet: The concentrations of mouse IFN-bweremeasured by enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, Cat#E-EL-M0033)according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Transfection, Control, Knockdown, Plasmid Preparation, Infection, Western Blot, Virus, Fluorescence, Microscopy, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Bilirubin metabolism in the liver orchestrates antiviral innate immunity in the body.

doi: 10.1016/j.celrep.2025.115481

Figure Lengend Snippet: Figure 7. UGT1A1 and UCB regulate IFN-I signaling in vivo (A) Mice were administered siCtrl or siUGT1A1 (66.7 mg/day, i.v.) for 72 h and then infected with VSV (1 3 108 PFU per gram body mouse, i.p.) for 24 h. (B–D) RT-qPCR analysis of Ugt1a1 mRNA (B), Ifnb mRNA (C), and VSV RNA (D) in the liver tissues from mice treated as in (A). (E) ELISA analysis of mIFNb protein levels in the liver tissues from mice treated as (A). (F) RT-qPCR analysis of the representative ISGs (Ifit1, Viperin, and Isg15) in the liver tissues from mice treated as in (A). (G) The mice were administered UCB or the vehicle (25 mg/kg body weight, i.p.) for 1 h and then given i.p. injections of mIFNb (10,000 IU per mouse) for 6 h. (H) RT-qPCR analysis of the representative ISGs (Ifit1, Viperin, and Mx1) in the liver tissues from mice treated as in (G). (I and J) RT-qPCR analysis of the representative ISG (Ifit1) in the kidney (I) and heart (J) tissues from mice treated as in (G).

Article Snippet: The concentrations of mouse IFN-bweremeasured by enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, Cat#E-EL-M0033)according to the manufacturer’s instructions.

Techniques: In Vivo, Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Biomaterials

Article Title: The antiviral activity of poly-γ-glutamic acid, a polypeptide secreted by Bacillus sp., through induction of CD14-dependent type I interferon responses.

doi: 10.1016/j.biomaterials.2013.08.067

Figure Lengend Snippet: Fig. 2. Induction of type I IFN responses by high-molecular weight g-PGAs. (A) HEK293hTLR4 cells grown in 6-well plates were transfected with plasmids expressing MD2 and CD14 (500 ng each/well) and with the luciferase expressing plasmids for the IFN-b promoter activity assay as in Fig. 1A. The transfected cells were treated with 100 mM g-PGAs with different molecular weights. As positive controls for the assay, the indicated concentrations of LPS and 0.2 mg/ml poly(I:C) were used to stimulate IFN-b promoter activity. Data are presented as means SD of three measurements from three independent experiments. *P < 0.05. (BeD) HEK293hTLR4 cells were transfected with plasmids expressing MD2 and CD14 and exposed to 100 mM g-PGAs with different molecular weights, 200 ng/ml LPS, or 0.1 mg/ml poly(I:C) for 30 h and then harvested for analysis of the secreted IFN-b levels by ELISA (B), and the mRNA levels of the type I IFN-inducible genes 20e50 OAS (C) and ISG56 (D) by real-time qRT-PCR. The GAPDH-normalized mRNA level in the mock-treated cells is considered one unit and the increase in mRNA abundance is shown as fold induction. ND ¼ not detected. The data shown are from one representative of duplicate experiments with triplicate assays. The two experiments gave highly similar results.

Article Snippet: The amounts of IFN-b secreted from the stimulated cells were assessed using the VeriKine human or mouse IFN-b enzyme-linked immunosorbent assay (ELISA) kit (PBL Interferon Source, Piscataway, NJ, USA).

Techniques: High Molecular Weight, Transfection, Expressing, Luciferase, Activity Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Biomaterials

Article Title: The antiviral activity of poly-γ-glutamic acid, a polypeptide secreted by Bacillus sp., through induction of CD14-dependent type I interferon responses.

doi: 10.1016/j.biomaterials.2013.08.067

Figure Lengend Snippet: Fig. 3. Induction of IFN-b production by g-PGA in hPBMC and mouse macrophage cells. hPBMCs (A) or RAW 264.7 cells (B) were treated with g-PGA, LPS, or poly(I:C) as in Fig. 2A or transfected with HCV 30-UTR in vitro transcripts as in Fig. 1C, for 8 h or 24 h. Levels of secreted IFN-b were measured by ELISA. Data from one of two independent experiments with similar results are shown.

Article Snippet: The amounts of IFN-b secreted from the stimulated cells were assessed using the VeriKine human or mouse IFN-b enzyme-linked immunosorbent assay (ELISA) kit (PBL Interferon Source, Piscataway, NJ, USA).

Techniques: Transfection, In Vitro, Enzyme-linked Immunosorbent Assay