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: The IFNAR1 neutralizing antibody MAR1 or mouse IgG (HY- P99137 and HY-P99977 purchased from MedChem Express LLC) was delivered intra-ganglionically on 3 dpi, at a dose of 1 μL(10 μg/1 μL).

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: 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: The IFNAR1 neutralizing antibody MAR1 or mouse IgG (HY- P99137 and HY-P99977 purchased from MedChem Express LLC) was delivered intra-ganglionically on 3 dpi, at a dose of 1 μL(10 μg/1 μL).

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: 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: The IFNAR1 neutralizing antibody MAR1 or mouse IgG (HY- P99137 and HY-P99977 purchased from MedChem Express LLC) was delivered intra-ganglionically on 3 dpi, at a dose of 1 μL(10 μg/1 μL).

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

Journal: Cell Research

Article Title: Depression compromises antiviral innate immunity via the AVP-AHI1-Tyk2 axis

doi: 10.1038/s41422-022-00689-9

Figure Lengend Snippet: a Western blot analysis of Tyr701 phosphorylation of STAT1 (p-STAT1) in Ahi1 +/+ and Ahi1 −/− mouse peritoneal macrophages treated with mIFNβ (1000 IU/mL). b Western blot analysis of IFNAR1 and IFNAR2 protein levels in Ahi1 +/+ and Ahi1 −/− mouse peritoneal macrophages. c Western blot analysis of the JAK-STAT signaling proteins (JAK1, Tyk2, STAT1, STAT2 and IRF9) in Ahi1 +/+ and Ahi1 −/− mouse peritoneal macrophages. d Western blot analysis of the JAK-STAT signaling proteins (JAK1, Tyk2, STAT1, STAT2 and IRF9) in RAW264.7 cells transfected with either control shRNAs (shCtrl) or two specific shRNAs against AHI1 (shAHI1: #1, #2). e RT-qPCR analysis of Tyk2 mRNA levels in RAW264.7 cells transfected with either shCtrl or two specific shAHI1 (#1, #2). f Western blot analysis of Tyk2 protein levels in Ahi1 +/+ and Ahi1 −/− mouse peritoneal macrophages treated with cycloheximide (CHX, 50 μg/mL) for durations. g Western blot analysis of Tyk2 protein levels in the spleen tissues from normal control mice (Normal, n = 5) or depression model mice ( n = 5). The ratio values indicate Tyk2/β-actin densitometric ratios analyzed by the ImageJ program. h Western blot analysis of Tyk2 and STAT1 protein levels in human PBMCs from healthy controls (Healthy, n = 5) and MDD patients ( n = 5). ns, not significant ( P > 0.05) (two-tailed unpaired Student’s t -test). Data are shown as means ± SD of three biological replicates ( e , f ), or are representative of three independent experiments ( a – d , f – h ).

Article Snippet: The antibodies with indicated dilutions were as follows: AHI1 (Santa Cruz, sc-515382, 1:500), JAK1 (Santa Cruz, sc-1677, 1:1000), Tyk2 (Cell Signaling Technology, 14193, 1:1000), STAT1 (Cell Signaling Technology, 9172, 1:1000), STAT2 (Cell Signaling Technology, D9J7L, 1:1000), IRF9 (Abcam, ab51639, 1:1000), IFNAR1 (Sino Biological, 13222-H08H, 1:1000), IFNAR2 (Santa Cruz, sc-137209, 1:1000), OTUD1 (Abcam, ab122481, 1:1000), Flag (Sigma, F7425, 1:5000), HA (Abcam, ab9110, 1:3000), Myc (Abmart, M20002H, 1:3000), pY701-STAT1 (Cell Signaling Technology, 9167, 1:1000), VSV-G (Santa Cruz, sc-66180, 1:1000), Tyk2 (Affinity, AF5223, 1:1000), β-Actin (Proteintech, 66009-1-Ig, 1:2000), Tubulin (Proteintech, 66031-1-Ig, 1:3000), Ubiquitin (Ub) (Santa Cruz, sc-8017, 1:500), K48-Ub (Cell Signaling Technology, 8081, 1:1000).

Techniques: Western Blot, Transfection, Quantitative RT-PCR, Two Tailed Test

Journal: Biomolecules

Article Title: Antiviral Functions of Type I and Type III Interferons in the Olfactory Epithelium

doi: 10.3390/biom13121762

Figure Lengend Snippet: Upregulation of type I and III interferon transcript levels. Relative expression of Ifna2, Ifna4, Ifnb2, and Ifnl2/3 in the olfactory mucosa using qRT-PCR ( A – D ). Relative expression of Ifnar1 and Ifnlr1 in the olfactory mucosa at 24 h PI ( E ). Biological triplicates were included ( A – E ). Immunostaining of IFNAR1 (green in ( F )) and IFNLR1 (green in ( G )) with OMP (red in ( F , G )) in the OE. Bar = 15 μm.

Article Snippet: The antibodies used were: chicken anti-OMP (custom, 1:1000), goat anti-GFP (Rockland (Baltimore, MD, USA), 2.2 µg/mL), rabbit anti-IFNAR1 (Sino Biological (Beijing, China), 1:20), goat anti-IFNLR1 (Novus Biologicals (Centennial, CO, USA), 10 μg/mL), and rabbit anti-pSTAT1 (Abcam, 3.0 µg/mL).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining

Journal: Biomolecules

Article Title: Antiviral Functions of Type I and Type III Interferons in the Olfactory Epithelium

doi: 10.3390/biom13121762

Figure Lengend Snippet: Interferon signaling is required for suppressing VSV replication in the olfactory mucosa. The expression levels of the viral genes, VSV-GFP, VSV-M, and VSV-N, in olfactory mucosae at 24 h PI were measured in Ifnar1 −/− ( A ), Ifnlr1 −/− ( B ), Ifnar1 −/− ; Ifnlr1 −/− ( C ), and Stat1 −/− ( D ) and compared to wildtype littermates. Student t -test, * p < 0.05.

Article Snippet: The antibodies used were: chicken anti-OMP (custom, 1:1000), goat anti-GFP (Rockland (Baltimore, MD, USA), 2.2 µg/mL), rabbit anti-IFNAR1 (Sino Biological (Beijing, China), 1:20), goat anti-IFNLR1 (Novus Biologicals (Centennial, CO, USA), 10 μg/mL), and rabbit anti-pSTAT1 (Abcam, 3.0 µg/mL).

Techniques: Expressing

Journal: Oncogene

Article Title: Evaluating the therapeutic potential of ADAR1 inhibition for triple-negative breast cancer

doi: 10.1038/s41388-020-01515-5

Figure Lengend Snippet: A) Core ISG Score in TNBC and Non-TNBC breast cancer samples. Data were extracted from TCGA database. B) Core ISG Score in ER-positive, ERBB2(HER2)-positive and TNBC cell lines. Data were extracted from CCLE database. C) ADAR1-dependency score positively correlates with Core ISG Score in breast cancer cell lines. Upper panel: Core ISG Score in breast cancer cell lines. Lower panel: ADAR1-dependency scores. D) Immunoblots showing protein levels of IFNAR1, PKR, p-PKR (T446), p-eIF2α (S51) and GAPDH (loading control) in MDA-MB231 cells. IFNAR1 was knocked down in ShADAR1-treated MDA-MB231 cells to determine if IFNAR1 loss reverses ADAR1-knockdown phenotype. Images are representative, N=3. F) FF assay showing that IFNAR1 loss partially rescued ADAR1-knockdown phenotype in MDA-MB231 cells. Images are representative, N=3. G) Quantification of FF in F) . Relative plate occupancy was determined using ImageJ software and normalized to ShNT-ShNT. Data are represented as mean ± SD. N=3.

Article Snippet: Primary antibodies used in this study include ADAR1 (Santa Cruz, Dallas, TX USA, sc-73408; Bethyl Laboratories, Montgomery, TX USA, A303–883A; Abcam, Cambridge, MA USA, ab126745), cleaved PARP (Cell Signaling, #9541), PKR (Cell Signaling, #3072), PKR Thr-446-P (Abcam, ab32036), IFNAR1 (Bethyl, A304–290A), ISG15 (Santa Cruz, sc-166755), GAPDH (Bethyl, A300–641A), β-Tubulin (Abcam, ab6046), EIF2S1/eIF2α Ser-51-P (Abcam, 32157), EIF2S1 (Abcam, ab5369), ATF4 (Cell Signaling, #11815).

Techniques: Western Blot, Control, Knockdown, Software