tbk1 kinase enzyme system kit  (Promega)

Journal: Eye and Vision

Article Title: STING deficiency alleviates scar formation after glaucoma filtration surgery by suppressing p38 MAPK-induced inflammation in mice

doi: 10.1186/s40662-026-00475-3

Figure Lengend Snippet: In vivo effects of Sting gene knockout on inflammation and fibrosis after glaucoma filtration surgery (GFS). a Relative Sting mRNA expression in the Sham and Sting −/− groups (n = 5 eyes in each group). b Intraocular pressure (IOP) changes in the WT + GFS and Sting −/− + GFS groups over 28 days post-surgery (n = 10 eyes in each group). c Representative images of bleb morphology in the Sham, WT + GFS, and Sting −/− + GFS groups. Yellow circles indicate the blebs. d Bleb survival analysis in the WT + GFS and Sting −/− + GFS groups using Kaplan–Meier survival curves (n = 5 eyes in each group). e Hematoxylin and eosin (H&E) staining of tissue sections from the Sham, WT + GFS, and Sting −/− + GFS groups (n = 3 eyes in each group). The triangles indicate the conjunctiva, the stars indicate the retina. f Immunofluorescence staining of α-SMA in the Sham, WT + GFS, and Sting −/− + GFS groups (n = 3 eyes in each group). The triangles indicate the conjunctiva. g Masson staining of tissue sections from the Sham, WT + GFS, and Sting −/− + GFS groups. The triangles indicate the conjunctiva, the stars indicate the retina. h Sirius ed staining of tissue sections from the Sham, WT + GFS, and Sting −/− + GFS groups. The triangles indicate the conjunctiva, the stars indicate the retina. i Analysis of Masson staining (n = 3 eyes in each group). j Analysis of Sirius red staining (n = 3 eyes in each group). k Relative mRNA expression of fibronectin, CTGF, COL1A1, α-SMA, and COL3A1 in the Sham, WT + GFS, and Sting −/− + GFS groups (n = 5 eyes in each group). l , m Immunofluorescence staining of p-TBK1 and p-IRF3 in the Sham, WT + GFS, and Sting −/− + GFS groups (n = 3 eyes in each group). The triangles indicate the conjunctiva, the stars indicate the retina. Arrows indicate the p-TBK1 expression area. Results were expressed as mean ± SD except ( b ) that was expressed as median (lower to upper quartile values). Statistical analysis was performed using unpaired t -test with Welch’s correction in ( a ), Wilcoxon signed-ranks test with Bonferroni correction in ( b ), log-rank test in ( d ), and one-way ANOVA followed by Bonferroni’s post-hoc test in ( i , j , k ). * P < 0.05, ** P < 0.01, *** P < 0.001. Sham, sham-operated; WT, wild-type; α-SMA, α-smooth muscle actin; CTGF, connective tissue growth factor; COL1A1, collagen type I alpha 1; COL3A1, collagen type III alpha 1; p-TBK1, phospho-tank-binding kinase 1; p-IRF3, phospho-interferon regulatory factor 3

Article Snippet: The following antibodies were used for immunofluorescence and immunohistochemistry: anti-STING (13647, Cell Signaling Technology, USA), anti-α-SMA (14395-1-AP, ProteinTech, China), anti-phospho-tank-binding kinase 1 (TBK1) (5483, Cell Signaling Technology, USA), anti-phospho-interferon regulatory factor 3 (IRF3) (29047, Cell Signaling Technology, USA), anti-phospho-p65 (3033, Cell Signaling Technology, USA), and anti-F4/80 (R013726, Epizyme, China).

Techniques: In Vivo, Gene Knockout, Filtration, Expressing, Staining, Immunofluorescence, Binding Assay

Journal: Microbiology Spectrum

Article Title: African swine fever virus MGF505-3R facilitates ferroptosis to restrict TBK1-IRF3 pathway

doi: 10.1128/spectrum.03423-24

Figure Lengend Snippet: MGF505-3R blocks the TBK1-IRF3 phosphorylation induced by GPX4 activation to resist IFN-β expression. PK-15 cells were transfected with pcDNA3.1-MGF505-3R-Flag plasmid and treated with GW7647 or poly(dA:dT). Protein and RNA were extracted for analyzing the TBK1-IRF3 pathway and IFN-β mRNA expression. ( A ) GPX4 expression. ( B ) pIRF3 and IRF3 expression. ( C ) pTBK1 and TBK1 expression. ( D ) IFN-β mRNA expression. ( E ) ISG15 mRNA expression. ( F ) ISG54 mRNA expression. * P < 0.05, ** P < 0.01, ***P < 0.001. ns, not significant.

Article Snippet: Anti-IRF3 and TANK-binding kinase 1 (TBK1) polyclonal antibodies were purchased from CST, USA.

Techniques: Phospho-proteomics, Activation Assay, Expressing, Transfection, Plasmid Preparation

Journal: Microbiology Spectrum

Article Title: African swine fever virus MGF505-3R facilitates ferroptosis to restrict TBK1-IRF3 pathway

doi: 10.1128/spectrum.03423-24

Figure Lengend Snippet: A working model of MGF505-3R promoting ferroptosis to restrict type I IFN production. ASFV infection and MGF505-3R transfection facilitate iron and ROS accumulation, resulting in NCOA4-mediated ferritinophagy and ferroptosis, while the GPX4 and Keap1-Nrf2 pathway is restricted. GPX4 activation promotes the TBK1-IRF3-IFN-β pathway and exerts antivirus capacity.

Article Snippet: Anti-IRF3 and TANK-binding kinase 1 (TBK1) polyclonal antibodies were purchased from CST, USA.

Techniques: Infection, Transfection, Activation Assay