Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Natterin bridges IFN-φ1 and non-canonical inflammasome pathways via CRFB1 /Gbp4 to license Caspy2-mediated antibacterial immunity

doi: 10.3389/fcimb.2025.1686758

Figure Lengend Snippet: IFN-φ neutralization abolishes ST-induced interferon production in zebrafish embryos. (A) Representative immunohistochemical staining of IFN-φ (brown DAB precipitate) in 1 dpf wild-type (WT) embryos under four conditions: (a.1) unstimulated control, (a.2) ST stimulation (10 6 cells/ml for 2h), (a.3) anti-IFN-β antibody pretreatment alone, and (a.4) anti-IFN-β pretreatment followed by ST stimulation. Scale bar: 50 μm. (B) Quantification of IFN-φ-positive cells from two slides per group (eight sections/slide, three embryos/section) using Fiji-ImageJ. Data represent mean ± SEM ( n = 3 independent experiments); * p < 0.05 versus unstimulated control WT, # p < 0.05 versus ST-stimulated WT.

Article Snippet: Independent groups of 1 dpf embryos ( n = 60) were previously neutralized by immersion in fresh 0.5× E2 medium for 30 min with the monoclonal mouse IgG2a anti-human IFN-β neutralizing antibody (clone 13A2, #mabg 2 hifnb-3, Invivogen) at 2 μg/ml and then exposed or not by immersion to ST for 2h, according to .

Techniques: Neutralization, Immunohistochemical staining, Staining, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Natterin bridges IFN-φ1 and non-canonical inflammasome pathways via CRFB1 /Gbp4 to license Caspy2-mediated antibacterial immunity

doi: 10.3389/fcimb.2025.1686758

Figure Lengend Snippet: IFN-I signaling is essential for IL-1β–mediated protection against ST stimulation. (A) Locomotor activity (total distance traveled) assessed 72h after stimulation at 28°C. Mortality count (C) and Caspy2 expression in WT embryos after ST stimulation (10 6 cells/ml, 2h) with or without anti-IFN-β antibody pretreatment. The corresponding bar graph (D) represents the protein expression levels as a percentage of the control. (E) Quantification and (F) representative IHC images of mature IL-1β (mIL-1β)–positive cells (brown DAB precipitate) in: (f.1) ST-stimulated embryos, (f.2) pretreatment with anti-IFN-β alone, and (f.3) pretreatment with anti-IFN-β followed by ST stimulation. Scale bar: 200 μm. Data represent mean ± SEM ( n = 3 independent experiments); * p < 0.05 versus unstimulated control WT, # p < 0.05 versus ST-stimulated WT.

Article Snippet: Independent groups of 1 dpf embryos ( n = 60) were previously neutralized by immersion in fresh 0.5× E2 medium for 30 min with the monoclonal mouse IgG2a anti-human IFN-β neutralizing antibody (clone 13A2, #mabg 2 hifnb-3, Invivogen) at 2 μg/ml and then exposed or not by immersion to ST for 2h, according to .

Techniques: Activity Assay, Expressing, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Natterin bridges IFN-φ1 and non-canonical inflammasome pathways via CRFB1 /Gbp4 to license Caspy2-mediated antibacterial immunity

doi: 10.3389/fcimb.2025.1686758

Figure Lengend Snippet: Effectiveness of CRISPR/Cas9 depletion of loc795232. Basal expression of loc795232 mRNA transcripts in unstimulated WT or KO embryos ( n = 100/group) at 1 dpf by RT-qPCR (A) or by in situ hybridization (B) . All qPCR data normalized to β-actin and expressed as fold change relative to 0h WT unstimulated control. Data represent mean ± SEM of three biological replicates. Independent groups of 1 dpf embryos previously treated or not with Pam3CSK4 or MCC950 and subjected to ST stimulation for 2h, as well as KO larvae stimulated with ST were processed for Natterin detection by WB (C) using anti-Natterin serum (62 kDa dimeric form) and anti-IgG TrueBlot HRP. The corresponding bar graph represents the protein expression levels as a percentage of the control (D) , and Ponceau S staining is visualized in the first horizontal line.

Article Snippet: Independent groups of 1 dpf embryos ( n = 60) were previously neutralized by immersion in fresh 0.5× E2 medium for 30 min with the monoclonal mouse IgG2a anti-human IFN-β neutralizing antibody (clone 13A2, #mabg 2 hifnb-3, Invivogen) at 2 μg/ml and then exposed or not by immersion to ST for 2h, according to .

Techniques: CRISPR, Expressing, Quantitative RT-PCR, In Situ Hybridization, Control, Staining

Journal: Oncoimmunology

Article Title: Mild microwave hyperthermia promotes mitotic catastrophe, induces time-delayed cGAS-STING activation and restores sensitivity to anti-PDL1 therapy in Pan02 pancreatic cancer model

doi: 10.1080/2162402X.2025.2602216

Figure Lengend Snippet: (A–I) Representative images of cGAS, STING, pSTAT1, IFNβ, CCL5, CXCL10, HMGB1, PD-L1, and γH2AX immunofluorescent or immunohistochemical stained tissue sections, respectively, of untreated, αPD-L1 mAb-treated, Ht-treated and Ht+αPD-L1 mAb-treated tumors (treatment side and abscopal side).

Article Snippet: The following antibodies were used for western blotting and immunostaining: anti-human cGAS (Cell Signaling), anti-mouse cGAS (SantaCruz), STING (Novus Biological), phospho-IRF3 Ser-386 (Cell Signaling), phospho-IRF3 Ser-396 (Cell Signaling), IRF-3 (SantaCruz), phospho-TBK1 Ser172 (Cell Signaling), TBK1 (SantaCruz), phospho-H2A.X Ser139 (Cell Signaling), PD-L1 (Cell Signaling), CD11c (Cell Signaling), CD8α (Cell Signaling), F4/80 (Proteintech), CD163 (Proteintech), CD86 (Novus Biologicals), FoxP3 (Cell Signaling), Gr1 (Invitrogen), GranzymeB (Invitrogen), Tim3 (Invitrogen), phospho-STAT1 Tyr701 (Cell Signaling), and IFNβ, CCL5, CXCL10, CD56, MICA, HMGB1, SUMO1, SUMO2/3, UBC9, SAE1, UBA2, SENP3, Beta-Actin (all from Proteintech).

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Leukocyte Biology

Article Title: The macrocyclic peptide rhesus theta defensin 1 activates interferon and antiviral pathways in human monocytes

doi: 10.1093/jleuko/qiaf150

Figure Lengend Snippet: Stimulation of ISRE reporter by RTD-1. The activity of Lucia luciferase secreted in the medium was assayed and the luminescence units are plotted. (A) THP-1 Dual cells were stimulated with HOAc (vehicle) or increasing concentrations of RTD-1. Results are from 3 experiments. * P < 0.05, 2-tailed t test, comparison with HOAc-treated sample. Error bars indicate standard deviation. (B) THP-1 Dual cells were pretreated with DMSO or Ruxo as indicated. Assay done in triplicate. * P < 0.05 when compared with HOAc-treated control, 2-tailed t test. # P < 0.05 when compared with RTD-1 treatment. Error bars indicate standard deviation. (C) THP-1 Dual cells were transferred to RPMI + 1% HI FBS + P/S + normocin medium and either 2.5 μg human IgG1 or anti-IFNAR1 antibodies were added to the culture. Cells were incubated for 2 h, and RTD-1 or IFN-β was added as indicated and the incubation continued for 18 h. Assay was done in triplicate. * P < 0.05 when compared with HOAc-treated control, 2-tailed t test. Error bars indicate standard deviation. (D) THP-1 Dual cells were treated with RTD-1, IFN-β, or both as shown. Assay was done in triplicate. * P < 0.05, 2-tailed t test. Error bars indicate standard deviation.

Article Snippet: IFN-β was purchased from R&D Systems and human IgG1 and human anti IFNAR1 antibodies were purchased from Selleck USA.

Techniques: Activity Assay, Luciferase, Comparison, Standard Deviation, Control, Incubation

Journal: Journal of Leukocyte Biology

Article Title: The macrocyclic peptide rhesus theta defensin 1 activates interferon and antiviral pathways in human monocytes

doi: 10.1093/jleuko/qiaf150

Figure Lengend Snippet: ISRE stimulation activity is distinct from RTD-1–mediated inhibition of NF-κB activation. THP-1 Dual cells were treated with LPS, RTD-1, or IFN-β or vehicle as shown and the medium was assayed for ISRE reporter activity (top) and NF-κB reporter activity (bottom) using Quanti-Luc and Quanti-Blue assays, respectively. Assay done in triplicate. # P > 0.05 (nonsignificant) when compared with respect to LPS treated samples, 2-tailed t test. Error bars indicate standard deviation.

Article Snippet: IFN-β was purchased from R&D Systems and human IgG1 and human anti IFNAR1 antibodies were purchased from Selleck USA.

Techniques: Activity Assay, Inhibition, Activation Assay, Standard Deviation