Journal: Cell reports

Article Title: Modular Architecture of the STING C-Terminal Tail Allows Interferon and NF-κB Signaling Adaptation

doi: 10.1016/j.celrep.2019.03.098

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Western blot analysis was performed using the following antibodies: high affinity anti-hemagglutinin (Sigma, anti-HA-Peroxidase, rat mAb 3F10), rabbit anti-TBK1/NAK (Cell Signaling-3013), rabbit anti-TRAF6 (abcam-EP591Y ab33915), rabbit anti-STING (Cell Signaling-D2P2F), anti-RPS19 (Bethyl-A304-002A) and as secondary antibody anti-rabbit-HRP (GE Healthcare-NA934).

Techniques: Virus, Recombinant, Reporter Assay, Expressing, Plasmid Preparation, Construct, Software

Journal: EBioMedicine

Article Title: STING-mediated intestinal barrier dysfunction contributes to lethal sepsis

doi: 10.1016/j.ebiom.2019.02.055

Figure Lengend Snippet: Stimulator of interferon genes (STINGs) was activated in sepsis and correlates with intestinal inflammation and gut barrier dysfunction. (A) Representative images of intestinal histology (H&E staining) in control group and sepsis patients. (B) Expression level of STING and (C) NF-κB in human intestines were analyzed through IHC staining. (D) Activation of IRF3 signaling in gut of sepsis patients. (E) Correlation between apoptosis indexes and expression of STING in human gut. (F, G) Correlation of inflammatory cytokines and I-FABP to the expression level of STING. Image J was used to detect STING optical density, and each symbol represents an individual patient. H&E, hematoxylin and eosin; IHC, immunohistochemistry, IRF3, interferon regulatory factor 3; I-FABP, intestinal fatty acid binding protein. ⁎ P < .05, ⁎⁎ P < .01 vs control group.

Article Snippet: For immunohistochemistry (IHC), slides were treated with anti-STING antibody (D2P2F; 13,647; Cell Signaling), anti-NF-κB antibody (ab16502; Abcam) according to the manufacturer's instructions.

Techniques: Staining, Control, Expressing, Immunohistochemistry, Activation Assay, Binding Assay

Journal: EBioMedicine

Article Title: STING-mediated intestinal barrier dysfunction contributes to lethal sepsis

doi: 10.1016/j.ebiom.2019.02.055

Figure Lengend Snippet: Induction of STING signaling in CLP-induced sepsis. (A) H&E staining for intestinal histology after CLP treatment. (B) intestinal cytokines levels. (C, D) Activation of STING signaling in intestine following CLP was analyzed by IHC staining and western blot. CLP, cecum ligation perforation. Data are expressed as the mean ± SD. ⁎ P < .05 vs sham group; n = 6 mice per group.

Article Snippet: For immunohistochemistry (IHC), slides were treated with anti-STING antibody (D2P2F; 13,647; Cell Signaling), anti-NF-κB antibody (ab16502; Abcam) according to the manufacturer's instructions.

Techniques: Staining, Activation Assay, Immunohistochemistry, Western Blot, Ligation

Journal: EMBO Reports

Article Title: DSTYK phosphorylates STING at late endosomes to promote STING signaling

doi: 10.1038/s44319-025-00394-9

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit polyclonal anti-STING , ABclonal , A3575.

Techniques: Recombinant, Bioprocessing, Sequencing, shRNA, Transfection, Real-time Polymerase Chain Reaction, SYBR Green Assay, Mutagenesis, Software

Journal: Frontiers in Microbiology

Article Title: Porphyromonas gingivalis induces an inflammatory response via the cGAS-STING signaling pathway in a periodontitis mouse model

doi: 10.3389/fmicb.2023.1183415

Figure Lengend Snippet: Porphyromonas gingivalis activates the cGAS-STING signaling pathway in vitro . HGFs were infected with P. gingivalis at an MOI of 100 for 6 h. Untreated cells (MOI 0 for 0 h) were used as controls. (A) Total lysates were harvested, and cGAS and STING protein levels in HGFs were confirmed by Western blotting. β-Actin served as a control. (B) Transcript levels of cGAS and STING were measured by qRT–PCR and normalized to uninfected cells under each treatment using the threshold cycle (2 −ΔΔCt ) method with GAPDH as the reference gene. (C) Supernatants were collected, and the levels of IL-1β, IL-6, TNF-a, and IFN-β were analyzed by ELISA. (D) Cells were also collected to measure the gene expression of RANKL by qRT–PCR. (E) Western blotting analysis of RANKL expression in HGFs. Values are expressed as the mean ± SEM ( n = 4). The p value is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, no significance.

Article Snippet: The antibodies used for this assay were anti-cGAS antibody (1:250, Cat# A8335, ABclonal), anti-STING rabbit mAb (1:500, Cat# A21051, ABclonal), anti-RANKL rabbit mAb (1:500, Cat# A2550, ABclonal), anti-β actin antibody (1:10000, Cat# 380624, Zen Biotechnology), and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:1000, Cat# ab7090, Abcam).

Techniques: In Vitro, Infection, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Frontiers in Microbiology

Article Title: Porphyromonas gingivalis induces an inflammatory response via the cGAS-STING signaling pathway in a periodontitis mouse model

doi: 10.3389/fmicb.2023.1183415

Figure Lengend Snippet: The Porphyromonas gingivalis infection-driven cGAS-STING signaling pathway induces type I IFN gene expression in mouse models of periodontitis. The periodontitis model was established using C57BL/6 mice, and untreated animals served as the control group ( n = 4 mice per group). (A) Mouse gingival cells were preincubated with DCFH-DA (10 mM) to detect ROS-induced fluorescence. Statistical analysis of ROS-positive cells per field in the two groups. Uninfected mice were used as controls (scale bar, 1,000 μm). (B) Mouse gingival tissue lysates were collected to determine the levels of cGAS and STING by Western blotting. (C) Mouse gingival tissue was collected to measure the gene expression of cGAS and STING by qRT–PCR. (D) Serum samples were collected, and the levels of IL-6, IL-1β, TNF-a and IFN-β were analyzed by ELISA. (E,F) Mouse gingival tissues were also collected to measure the gene expression of IFN-β (E) and RANKL (F) by qRT–PCR. Values are expressed as the mean ± SEM. The p value is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, no significance.

Article Snippet: The antibodies used for this assay were anti-cGAS antibody (1:250, Cat# A8335, ABclonal), anti-STING rabbit mAb (1:500, Cat# A21051, ABclonal), anti-RANKL rabbit mAb (1:500, Cat# A2550, ABclonal), anti-β actin antibody (1:10000, Cat# 380624, Zen Biotechnology), and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:1000, Cat# ab7090, Abcam).

Techniques: Infection, Expressing, Fluorescence, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Microbiology

Article Title: Porphyromonas gingivalis induces an inflammatory response via the cGAS-STING signaling pathway in a periodontitis mouse model

doi: 10.3389/fmicb.2023.1183415

Figure Lengend Snippet: Porphyromonas gingivalis decreases proinflammatory cytokine synthesis in periodontitis Sting Gt mice via the cGAS-STING signaling pathway. Sting Gt and C57BL/6 mice were used to construct periodontitis models, with C57BL/6 periodontitis mice serving as controls ( n = 4 mice per group). (A) Diagram of the STING transgenic (Sting Gt ) mouse periodontitis model. (B) Images showing H&E staining following infection with P. gingivalis . The images shown are 14 dpi for all groups (scale bar, 200 μm). Each image is representative of a group of 4 mice at 14 dpi. Semiquantitative analysis of H&E pictures was also performed. (C) Serum samples were collected, and the levels of IL-6, IL-1β, TNF-a and IFN-β were analyzed by ELISA. (D) Mouse gingival tissues were also collected to measure the gene expression of IFN-β by qRT–PCR. (E–G) The gingival cells were harvested and stained with mAbs specific for F4/80, CD11c, CD206, CCR2, and CCL2 for flow cytometric analysis. The results are expressed as M1 macrophages: F4/80 + CD11c+; M2 macrophages: F4/80 + CD206+. The relative proportion of M1 macrophages and M2 macrophages is shown in (E) . The relative proportion of CCR2 (F) and CCL2 (G) in gingival cells induced by P. gingivalis . The results shown are 14 dpi for all groups. Data are expressed as the mean ± SEM. The p value is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, no significance.

Article Snippet: The antibodies used for this assay were anti-cGAS antibody (1:250, Cat# A8335, ABclonal), anti-STING rabbit mAb (1:500, Cat# A21051, ABclonal), anti-RANKL rabbit mAb (1:500, Cat# A2550, ABclonal), anti-β actin antibody (1:10000, Cat# 380624, Zen Biotechnology), and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:1000, Cat# ab7090, Abcam).

Techniques: Construct, Transgenic Assay, Staining, Infection, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Frontiers in Microbiology

Article Title: Porphyromonas gingivalis induces an inflammatory response via the cGAS-STING signaling pathway in a periodontitis mouse model

doi: 10.3389/fmicb.2023.1183415

Figure Lengend Snippet: Porphyromonas gingivalis decreases alveolar bone loss in periodontitis Sting Gt mice via the cGAS-STING signaling pathway. (A) Three-dimensional reconstruction images and analysis of the cementoenamel junction to alveolar bone crest (CEJ-ABC) distances of maxilla with periodontitis mice (scale bar: 500 μm). Representative X-ray images (scale bar: 1.0 mm) (B) Mouse periodontal tissues were also collected to measure the gene expression of RANKL by qRT–PCR.

Article Snippet: The antibodies used for this assay were anti-cGAS antibody (1:250, Cat# A8335, ABclonal), anti-STING rabbit mAb (1:500, Cat# A21051, ABclonal), anti-RANKL rabbit mAb (1:500, Cat# A2550, ABclonal), anti-β actin antibody (1:10000, Cat# 380624, Zen Biotechnology), and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:1000, Cat# ab7090, Abcam).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Microbiology

Article Title: Porphyromonas gingivalis induces an inflammatory response via the cGAS-STING signaling pathway in a periodontitis mouse model

doi: 10.3389/fmicb.2023.1183415

Figure Lengend Snippet: Suppression of cGAS/STING ameliorates Porphyromonas gingivalis -induced periodontal disease. P. gingivalis -induced periodontitis mouse groups were injected intraperitoneally with SR-717 (30 mg/kg), SN-011 (10 mg/kg) and vehicle (5% DMSO in PBS) for 7 days of daily dosing and euthanized after the last treatment. Mice with PBS-treated periodontitis served as controls ( n = 4 mice per group). (A) Mouse gingival tissue lysates were collected to determine the protein levels of cGAS and STING by Western blotting. (B) Serum samples were collected, and the levels of IL-6, IL-1β, TNF-a and IFN-β were analyzed by ELISA. (C) H&E staining showed hemi-maxillae histopathological changes after treatment with small molecule modulators (scale bar, 200 μm). Semiquantitative analysis of H&E pictures was also performed. (D–F) Gingival cells were harvested and stained with mAbs specific for F4/80, CD11c, CD206, CCR2, and CCL2 for flow cytometric analysis. The results are expressed as M1 macrophages: F4/80 + CD11c+; M2 macrophages: F4/80 + CD206+. The relative proportion of M1 macrophages and M2 macrophages is shown in (D) . The relative proportion of CCR2 (E) and CCL2 (F) in periodontal ligament cells induced by P. gingivalis . (G) Mouse gingival tissues were also collected to measure the gene expression of RANKL by qRT–PCR. Data are expressed as the mean ± SEM. The p value is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, no significance.

Article Snippet: The antibodies used for this assay were anti-cGAS antibody (1:250, Cat# A8335, ABclonal), anti-STING rabbit mAb (1:500, Cat# A21051, ABclonal), anti-RANKL rabbit mAb (1:500, Cat# A2550, ABclonal), anti-β actin antibody (1:10000, Cat# 380624, Zen Biotechnology), and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:1000, Cat# ab7090, Abcam).

Techniques: Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Quantitative RT-PCR