Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet: Glab inhibits the activation of the cGAS-STING signaling pathway. A Glabridin (Glab) structure. B and C BMDMs were first treated with DMSO or Glab of various concentrations for 1 h, and then stimulated with HT-DNA or Poly(I:C) for 2 h. Whole cell lysate (WCL) was collected and immunoblotted with the indicated antibody. D – F BMDMs were treated with DMSO or Glab of various concentrations for 1 h, and then stimulated with HT-DNA for 4 h. The expression of IFN-β, IL-6 and TNF-α mRNA level were detected by quantitative polymerase chain reaction (qPCR) assay. G – I BMDMs were treated with DMSO or Glab of various concentrations for 1 h and then stimulated with Poly(I:C) for 4 h. The expression of IFN-β, IL-6 and TNF-α mRNA were detected by qPCR assay. Data in D – I are presented as mean ± SEM from three biological replicates. one-way ANOVA and Dunnett’s post hoc test were used to assess the differences of multiple groups, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control, NS, not significant

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet: Glab is a specific inhibitor of the cGAS-STING signaling pathway. A BMDMs were first treated with DMSO or Glab (20 μM) for 1 h, then stimulated with HT-DNA, Poly(I:C), cGAMP, DMXAA and diABZI to analyze the phosphorylation of IRF3 and the expression of STING in whole cell lysates (WCL) by immunoblotting. B Human PBMCs were first treated with DMSO or Glab (20 μM) for 1 h, and then stimulated with Poly(I:C), cGAMP and diABZI for 2 h. Whole cell lysate was collected and immunoblotted with the indicated antibody. ( C–E ) BMDMs were first treated with Glab (20 μM) for 1 h and then stimulated with HT-DNA, Poly(I:C), cGAMP, DMXAA and diABZI for 4 h. The expression of IFN-β, IL-6 and TNF-α mRNA were detected by qPCR assay. F – H Human PBMCs were first treated with Glab (20 μM) for 1 h and then stimulated with HT-DNA, Poly(I:C), cGAMP and diABZI for 4 h. The expression of IFN-β, IL-6 and TNF-α mRNA were detected by qPCR assay. Data in ( C–H ) are expressed as mean ± SEM (n = 3/group, from three biological replicates.). Unpaired student t -test C–H was used to assess differences among groups, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control, NS, not significant

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Expressing, Western Blot, Control

Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet: Glab inhibits the activation of STING and downstream signaling pathway in vivo. A – C Determination of IFN-β ( A ), IL-6 ( B ) and TNF-α ( C ) concentrations in collected serum by ELISA kits (n = 6 mice per group). D – F Determination of IFN-β ( D ), IL-6 ( E ) and TNF-α ( F ) concentrations in collected peritoneal lavage fluid by ELISA kits (n = 6 mice per group). G – I Cells were collected by centrifugation of peritoneal lavage fluid and analyzed for mRNA expression of relevant genes by qPCR assay (n = 6 mice per group). Data in ( A–I ) are expressed as mean ± SEM (n = 6 mice per group). one-way ANOVA and Dunnett's post hoc test were used to assess differences among groups, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control, NS, not significant

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Activation Assay, In Vivo, Enzyme-linked Immunosorbent Assay, Centrifugation, Expressing, Control

Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet: Glab inhibits the activation of the cGAS-STING pathway by disrupting the STING-IRF3 interaction. A Transfection of Flag-tagged plasmids (Flag-MAVS, Flag-STING, Flag-TBK1 and Flag-IRF3) into HEK-293 cells for 12 h, and then treated with vehicle, Glab (20 μM) for 6 h. Whole cell lysates were collected and immunoblotted with the indicated antibody. Samples for qPCR were detected by qPCR assay for the expression of IFN-β mRNA. B BMDMs were first treated with DMSO or Glab (20 μM) for 1 h and then stimulated with cGAMP for 2 h. The expression of STING and the oligomerization of SITNG in the cell lysate were analyzed by Western blot with the indicated antibodies. C and D HEK-293 T cells were transfected with Flag-tagged plasmids (Flag-Vector, Flag-IRF3 and Flag-TBK1) and HA-tagged plasmids (HA-Vector and HA-STING) for 20 h, then treated with Glab (20 μM) for 6 h and immunoprecipitated with Anti-FLAG® M2 Affinity Gel, as shown by Western Blots analysis. Data in A are expressed as mean ± SEM (n = 3/group, from three biological replicates.). one-way ANOVA and Dunnett’s post hoc test were used to assess differences among groups, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control, NS, not significant

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Activation Assay, Transfection, Expressing, Western Blot, Plasmid Preparation, Immunoprecipitation, Control

Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet:

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Transfection, SYBR Green Assay

Journal: Molecular Medicine

Article Title: Glabridin improves autoimmune disease in Trex1-deficient mice by reducing type I interferon production

doi: 10.1186/s10020-023-00754-y

Figure Lengend Snippet:

Article Snippet: Mouse IFN-β ELISA kit , Invivogen , luex-mifnbv2.

Techniques: Sequencing