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

Journal: Acta Pharmaceutica Sinica. B

Article Title: Proteomics and metabolic phenotyping define principal roles for the aryl hydrocarbon receptor in mouse liver

doi: 10.1016/j.apsb.2021.10.014

Figure Lengend Snippet: Metabolic phenotyping. Body composition was determined by (A) % change in final total body weight relative to the initial total body weight; and using dual energy X-ray absorptiometry scan to obtain measurements for (B) % body fat; and (C) % lean body mass. (D) Fasting plasma glucose levels was determined by Piccolo Xpress chemical analyzer; and (E) Plasma insulin level was measured by Luminex® 100 system. A glucose tolerance test (GTT) was performed and (F) curves for GTT were plotted; in addition, (G) the area under curve (AUC) was determined. Circulating lipids, namely, fasting plasma (H) total cholesterol; (I) HDL cholesterol; and (J) triglycerides were measured using Piccolo Xpress chemical analyzer. Values are mean ± SD. A complete list of P -values (determined by two-way ANOVA with Tukey's post-test) is provided in the accompanying table. In the figure panels, statistical significance is denoted by: a = genotype effect; b = PCB effect; c = interaction effect; d = WT Vehicle vs. WT PCB126; e = Ahr –/– Vehicle vs. Ahr –/– PCB126; f = WT Vehicle vs. Ahr –/– Vehicle; g = WT PCB126 vs. Ahr –/– PCB126. Ahr, aryl hydrocarbon receptor; PCB, polychlorinated biphenyl; WT, wild type.

Article Snippet: Plasma alanine transaminase (ALT), aspartate transaminase (AST), cholesterol, triglyceride, high-density lipoprotein (HDL), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), and non-HDL cholesterol (nHDLc) levels were determined with lipid panel plus kits on a Piccolo Xpress Chemistry Analyzer (Abbott Laboratories, Chicago, IL, USA).

Techniques: Clinical Proteomics, Luminex

Journal: Acta Pharmaceutica Sinica. B

Article Title: Proteomics and metabolic phenotyping define principal roles for the aryl hydrocarbon receptor in mouse liver

doi: 10.1016/j.apsb.2021.10.014

Figure Lengend Snippet: Characterization of fatty liver disease. Histological analysis of liver sections was performed using (A) hematoxylin-eosin (H&E) stain (10× magnification) and (B) Oil red O stain (10× magnification). The inset is 40× magnification. Hepatic lipids were extracted and measured including (C) liver triglycerides; (D) liver free fatty acids (FFA); and (E) liver cholesterol. Whole liver was isolated at euthanasia and (F) liver weight to total body weight ratio was calculated. Activity of circulating liver enzymes, namely (G) alanine aminotransferase (ALT); and (H) aspartate aminotransferase (AST) were calculated using Piccolo Xpress chemical analyzer. RT-PCR was performed to measure hepatic mRNA expression levels of (I) Cd36 ; and (J) Pnpla3 . Expression of mRNA was normalized to the WT Vehicle group. Values are mean ± SD. A complete list of P -values (determined by two-way ANOVA with Tukey's post-test) is provided in the accompanying table. In the figure panels, statistical significance is denoted by: a = genotype effect; b = PCB effect; c = interaction effect; d = WT Vehicle vs. WT PCB126; e = Ahr –/– Vehicle vs. Ahr –/– PCB126; f = WT Vehicle vs. Ahr –/– Vehicle; g = WT PCB126 vs. Ahr –/– PCB126. AHR, aryl hydrocarbon receptor; CD36, cluster of differentiation 36; PCB, polychlorinated biphenyl; PNPLA3, patatin-like phospholipase domain-containing protein 3; WT, wild type.

Article Snippet: Plasma alanine transaminase (ALT), aspartate transaminase (AST), cholesterol, triglyceride, high-density lipoprotein (HDL), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), and non-HDL cholesterol (nHDLc) levels were determined with lipid panel plus kits on a Piccolo Xpress Chemistry Analyzer (Abbott Laboratories, Chicago, IL, USA).

Techniques: Staining, Isolation, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Expressing