recombinant mouse ifn β  (Cell Signaling Technology Inc)

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Expression of IRF7 and IFNβ in the TG during CFA-induced inflammatory pain. ( A ) Quantification of IFNβ levels in the TG and serum by ELISA before and after injection of CFA at 6 h, 1 d, 3 d post-CFA injection.( B ) Quantitative analysis of mRNA expression of Ifnb and Irf7 in the TG at 6 h, 1, 3 and 7 d post-CFA injection. ( C ) Time course of IRF7 and IFNβ expression in the TG during orofacial inflammatory pain and quantitative analysis of their expression levels normalized to GAPDH. ( D ) Quantitative analysis of mRNA expression of the key upstream factors (Sting1, Tlr4, Tlr3 and Irf3) related to IFNβ synthesis. Data are expressed as mean ± SD; N= 3-5 per group; one-way ANOVA followed by post hoc test was performed to assess significance between groups (*P < 0.05, **P < 0.01)

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Injection

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Distribution of IRF7-IFNβ signaling in the wild-type TG. ( A , C , E ) Representative images and quantitative co-localization analysis of IRF7, IFNβ or IFNAR co-labeled with neuronal (NeuN), macrophage (CD68), and SGC (GS) markers in the TG (A,C scale bar = 100 μm; E, scale bar = 50 μm).( B , D ) Immunofluorescence images and quantitative co-localization analysis of IRF7 or IFNβ co-labeled with neuronal subtypes expressing CGRP, P2X3R, or NF in the TG (scale bar = 100 μm). ( F ) t-SNE plots showing the distribution of Irf7, Ifnar1, Ifnar2 and the markers of neuronal subtypes in TG neurons, including Calca (encode CGRP), P2rx3 (encode P2X3R) and Nefh (encode NF), based on single-cell sequencing. ( G ) Proportion of Irf7-positive cells co-expressing Nefh, Calca and P2rx3. ( H ) The distribution of Ifnar1 and Ifnar2 in various TG cell types

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Labeling, Immunofluorescence, Expressing, Single Cell, Sequencing

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Effects of silencing IRF7 on inflammatory pain. ( A ) Western blot analysis of IRF7 and IFNβ expression in the TG 4 weeks after intra-ganglionic injection of an AAV vector carrying either Irf7 shRNA (sh-Irf7) or scramble shRNA (sh-Scr) (N = 3). ( B ) Representative images of enhanced green fluorescent protein (eGFP, green) and immunofluorescence staining of IRF7 (red) in the TG after intra-ganglionic injection of saline (Ctrl), Irf7 shRNA or scramble shRNA (scale bar = 40 μm). The arrows highlight the co localization signals of IRF7 and eGFP (N = 4). ( C ) Mechanical hypersensitivity was measured using Von Frey filaments in mice injected with saline, Irf7 shRNA or scramble shRNA (N = 10). ( D ) Immunofluorescence staining of c-Fos (red) expression in the SpVc of mice from the Ctrl, sh-Irf7 and sh-Scr group (scale bar = 200 μm, N = 4). All data are expressed as mean ± SD; N = 6-10, *P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA followed by post hoc test

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Western Blot, Expressing, Injection, Plasmid Preparation, shRNA, Immunofluorescence, Staining, Saline

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Activation of IFNβ signaling contributes to mechanical nociceptive hypersensitivity in mice. ( A ) Schematic illustration of the experimental procedure of Fig. 5 B and C. ( B ) Mechanical hypersensitivity measured by Von Frey filaments in mice intraperitoneally injected with either PBS or IFNβ (10 4 /10 5 U) on day 3 following CFA injection (N = 6). ( C ) The mechanical pain threshold was assessed daily in CFA injected mice using von Frey test, following treatment with either IFNβ (10⁴ U) or PBS (N = 6). ( D - E ) Mechanical hypersensitivity was evaluated by Von Frey test in mice intra-ganglionic injected with an IFNAR1 antagonist (MAR1) or IgG 3 d post-CFA injection. ( F ) Mechanical hypersensitivity of mice receiving direct injection of IFNβ (20 μL) ( G - H ) Representative images of immunofluorescence staining showing c-Fos (red) in the SpVc of mice treated with PBS or IFNβ (N = 5), or mice receiving intra-ganglionically injection of IgG or MAR1(N = 3). (scale bar = 200 μm). ( I - J ) Quantitative analysis of c-Fos (red) expression in the SpVc among groups. Data are expressed as mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001; ( B , E ) using one-way ANOVA followed by post hoc test, while ( C , F , I and J ) using unpaired t test

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Activation Assay, Injection, Immunofluorescence, Staining, Expressing

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: IRF7 contributes to inflammatory pain through upregulating IFNβ expression in the TG. ( A ) Screenshot from the UCSC Genome Browser showing the predicted IRF7 binding sites on human IFNB1 gene using the UCSC database. ( B ) JASPAR output displaying predicted IRF7 binding sites and corresponding motifs on IFNB1. ( C ) Immunohistochemical staining of IRF7 and IFNβ in paired mirror-image tissue sections, scale bar = 100 μm, and corresponding magnified views. The arrows indicate positive signals of IRF7 and IFNβ in the same cell. The intersection of positive signals labeled IRF7 and IFNβ and their co-localization analysis of gray-scale signal intensities ( D ). ( E ) Co-immunoprecipitation evaluation of IRF7’s effect on IFNβ protein in the TG from mice injected with CFA at 3 dpi. ( F ) Mechanical allodynia measured by Von Frey test on mice received either Irf7 shRNA or scramble shRNA TG-injection followed by intraperitoneally administered with IFNβ or PBS under inflammatory pain conditions (Data are expressed as mean ± SEM; N = 10, *P < 0.05, **P < 0.01, compared to the mice injected with sh-Irf7 and PBS; #P < 0.05, ##P < 0.01, compared to the mice injected with sh-Scr and IFNβ; one-way ANOVA followed by post hoc test). ( J ) ( G ) Immunofluorescence staining of c-fos (red) in the SpVc of IRF7-knockdown mice treated with IFNβ or PBS under inflammatory pain conditions (scale bar = 200 μm, N = 4, *P < 0.05, unpaired t test)

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Expressing, Binding Assay, Immunohistochemical staining, Staining, Labeling, Immunoprecipitation, Injection, shRNA, Immunofluorescence, Knockdown

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Effects of IRF7-IFNβ signaling on neuronal sensitization and neuroinflammation. ( A ) Detection of cellular calcium changes over time in neurons stimulated with different concentrations of IFNβ. ( B ) Representative fluorescence images showing the expression of Substance P (SP) in neurons following IFNβ stimulation (scale bar = 40 μm). ( C ) mRNA expression levels of CGRP, P2XR, SP, and TRPV1 in mouse TG neurons stimulated with 100 U or 300 U IFNβ, or pretreated with IgG1κ/MAR1 overnight followed by 100 U IFNβ stimulation. ( D - E ) Representative images and quantitative analysis of GFAP and CD86 in the TG of CFA-treated mice after IFNβ treatment (scale bar = 100 μm). ( F ) Western blot analysis of IRF7, NLRP3, IL6 and GAPDH expression in the TG of mice treated with IFNβ or PBS under inflammatory pain conditions. ( G ) Western blot analysis of IRF7, NLRP3, IL6 and GAPDH expression in the TG of CFA- treated mice intra-ganglionically injected with IgG or an IFNAR1 antagonist. ( H ) Western blot analysis of the expression of IRF7, NLRP3, IL6 and GAPDH in the TG of CFA-injected mice receiving sh-Irf7 or sh-Scr stereotaxic microinjection. Data are expressed as mean ± SD, N = 3-7 mice. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, unpaired t test

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: Fluorescence, Expressing, Western Blot, Injection, Microinjection

Journal: Inflammation

Article Title: IRF7 Modulates Inflammatory Pain Through Upregulating IFNβ in Mice Trigeminal Ganglion

doi: 10.1007/s10753-026-02483-w

Figure Lengend Snippet: Schematic of the effect of IRF7-IFNβ signal in the TG during orofacial inflammatory pain. Our work demonstrates a pro-nociceptive effect of IRF7 in the TG using AAV-mediated shRNA transduction. The effect of IRF7 on pain modulation may depend on IFNβ, which induces TG neuroinflammation and pro-inflammatory cytokines production through autocrine or paracrine signaling

Article Snippet: Recombinant mouse IFNβ (8234-MB-010/CF, R&D Systems) was dissolved in phosphate buffered saline (PBS) and administered intraperitoneally (i.p.) at a final concentration of 10 4 or 10 5 U, in a volume of 0.25 mL per injection, at 7 days post CFA injection (dpi).

Techniques: shRNA, Transduction

Journal: Emerging Microbes & Infections

Article Title: TOR3A represses type I interferon production and limits viral clearance during respiratory syncytial virus infection

doi: 10.1080/22221751.2026.2637961

Figure Lengend Snippet: The upregulation of TOR3A during RSV infection is facilitated by STAT1. ( A ) Flowchart overview of the infection of RAW264.7 cells with RSV followed by mass spectrometry analysis. RAW264.7 cells were infected with RSV (MOI = 1) for 12 h, with 2% DMEM (RSV solvent) as control. ( B ) Whole protein profiles of RAW264.7 cells after 12-hour RSV infection were analyzed. Cluster analysis identified differentially expressed interferon-stimulated genes (ISGs) proteins between RSV-infected and control groups. ( C ) KEGG analysis demonstrated activation of multiple infection-related signalling pathways following RSV infection. ( D ) GO enrichment analysis showing significant activation of cellular functions in response to RSV infection. ( E - F ) RT-qPCR analysis of Ifnb and Tor3a mRNA levels in RAW264.7 cells at 0, 6, 12, and 24 h post-RSV infection. ( G ) Western blot analysis of TOR3A protein expression in RAW264.7 cells at indicated time points (0, 12, 24 h) post-RSV infection. ( H ) RT-qPCR analysis of TOR3A mRNA levels in PBMCs from hospitalized RSV-bronchiolitis patients (n = 70) and controls (n = 40). ( I ) RT-qPCR analysis of TOR3A expression in PBMCs from RSV-infected children with mild (n = 48) or moderate to severe (n = 22) disease. ( J ) Analysis of the correlation between TOR3A mRNA levels and RSV viral copy number. ( K ) ROC analysis was performed to evaluate the predictive value of TOR3A expression for bronchiolitis in RSV-infected children. ( L ) Analysis of TOR3A mRNA levels from peripheral blood transcriptomic data ( GSE188424 ) in hospitalized RSV-infected children (n = 24) versus age-matched healthy controls (n = 24). ( M ) ROC curve analysis of the predictive value of TOR3A for RSV infection. ( N-O ) RAW264.7 cells were stimulated with IFN-β at a working concentration of 1000 U/ml. The mRNA and protein levels of TOR3A were measured by RT-qPCR and Western blot, respectively. ( P-Q ) RAW264.7 cells were pre-incubated with an IFNAR1-specific blocking antibody (100 ng/mL) for 2 h prior to RSV infection (12 h). The mRNA and protein levels of TOR3A were then measured by RT-qPCR and Western blot, respectively. ( R ) ChIP analysis revealed that STAT1 binds to the Tor3a promoter. ( S ) HEK293 T cells were co-transfected for 24 h with a luciferase reporter plasmid driven by the TOR3A promoter, a STAT1 expression plasmid, and empty vector, followed by a dual-luciferase reporter assay. Data are representative of three independent experiments and presented as mean ± SD. Statistical significance was determined by Student's t-test (H, I, L, and S), one-way ANOVA followed by Dunett's multiple comparisons test (E, F, N, and P). ns: no significance, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Innate immune stimuli included poly(I:C) (Merck), 5′-ppp hairpin RNA, and the STING agonist DMXAA (both from InvivoGen); recombinant mouse IFN-β and IFN-λ2 was from R&D Systems.

Techniques: Infection, Mass Spectrometry, Solvent, Control, Activation Assay, Quantitative RT-PCR, Western Blot, Expressing, Concentration Assay, Incubation, Blocking Assay, Transfection, Luciferase, Plasmid Preparation, Reporter Assay

Journal: Emerging Microbes & Infections

Article Title: TOR3A represses type I interferon production and limits viral clearance during respiratory syncytial virus infection

doi: 10.1080/22221751.2026.2637961

Figure Lengend Snippet: TOR3A deficiency augments IFN-I production and restrains virus replication in vivo . ( A ) Establishment of an intranasal RSV infection model in mice. ( B ) ELISA of serum IFN-β in WT versus TOR3A KO mice post-RSV infection (1 × 10⁶ PFU, i.n.). ( C-D ) RT-qPCR analysis of RSV-F and RSV-M2-1 mRNA levels in the lungs of RSV-infected WT and TOR3A KO mice (n = 3 per group). ( E ) Plaque assay of RSV titres in lung homogenate supernatants from RSV-infected WT and TOR3A KO mice. ( F ) Western blot analysis of M2-1 protein expression in lung tissues from RSV-infected WT and TOR3A KO mice. ( G ) H&E staining and pathological scoring of lung sections from PBS- or RSV-treated WT and TOR3A KO mice. Scale bar, 100 μm. ( H ) Establishment of a VSV intraperitoneal infection model in mice. ( I ) RT-qPCR analysis of VSV load in the lungs of WT and TOR3A KO mice (n = 3 per group) after infection (4 × 10⁸ PFU). ( J ) Plaque assay of VSV titres in lung homogenate supernatants from VSV-infected WT and TOR3A KO mice. ( K ) Western blot analysis of VSV-G protein expression in lung tissues from VSV-infected WT and TOR3A KO mice. (L) H&E staining and histopathological scoring of lung sections from PBS- or VSV-treated WT and TOR3A KO mice. Scale bar, 100 μm. (M) Survival of WT and TOR3A KO mice (n = 20 per group) after intraperitoneal infection with VSV (4 × 10⁸ PFU). Data are representative of three independent experiments and presented as mean ± SD. Statistical significance was determined by Student's t-test (B, C, D, E, G, I, J, and K) or survival curve (L) followed by Log-rank (Mantel-Cox) test. ns: no significance, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Innate immune stimuli included poly(I:C) (Merck), 5′-ppp hairpin RNA, and the STING agonist DMXAA (both from InvivoGen); recombinant mouse IFN-β and IFN-λ2 was from R&D Systems.

Techniques: Virus, In Vivo, Infection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Plaque Assay, Western Blot, Expressing, Staining

Journal: bioRxiv

Article Title: The amplitude of gammaherpesvirus lytic replication dictates adaptive immune activation: Potential implications for KSHV LANA in immune evasion

doi: 10.64898/2026.02.05.703987

Figure Lengend Snippet: (A) Ifnar1 -deficient or non-target control (mSAFE) 3T12 fibroblast pools were stained with Ifnar1 -specific antibodies and analyzed by flow cytometry. (B) Cells were treated with recombinant IFN-β at indicated doses to determine efficacy of Ifnar1 -specific deletion. Lysates were collected 2.5 hours after treatment and immunoblot analyses were performed using antibodies to detect indicated proteins. (C-D,F) Cells were infected with the indicated virus at an MOI of 0.1 PFU/cell. (C) Viral titers were determined at the indicated times postinfection by plaque assay. (D) RNA was collected at the indicated times postinfection. Viral transcripts were quantified by qRT-PCR relative to B2m using the ΔΔCT method and normalized to transcript abundance at 24 hours postinfection. (E) Cells were infected at an MOI of 1 PFU/cell and lysates were collected 48 hours postinfection. Immunoblot analyses were performed using antibodies that recognize the indicated proteins. (F) Cells were fixed 72 hours postinfection and stained for indirect immunofluorescence analysis using antibodies that recognize the indicated proteins. DNA was stained with DAPI. Scale bar – 200um.

Article Snippet: Puromycin selected 3T12s were tested for Ifnar1 deletion by treatment with recombinant mouse IFN-β (Cell Signaling Technologies).

Techniques: Control, Staining, Flow Cytometry, Recombinant, Western Blot, Infection, Virus, Plaque Assay, Quantitative RT-PCR, Immunofluorescence