Journal: Frontiers in Immunology

Article Title: New MoDC-Targeting TNF Fusion Proteins Enhance Cyclic Di-GMP Vaccine Adjuvanticity in Middle-Aged and Aged Mice

doi: 10.3389/fimmu.2020.01674

Figure Lengend Snippet:

Article Snippet: Cyclic di-GMP (vaccine-grade) , Invivogen , Cat# vac-nacdg.

Techniques: Enzyme-linked Immunosorbent Assay, Protein Extraction, Lysis, Software

Journal: Journal of translational medicine

Article Title: Photothermal therapy combined with a STING agonist induces pyroptosis, and gasdermin D could be a new biomarker for guiding the treatment of pancreatic cancer.

doi: 10.1186/s12967-025-06247-2

Figure Lengend Snippet: Scheme 1 A schematic summary of this study, showing that PTT induces pyroptosis via caspase-1/GSDMD pathway and arouses antitumour immune response. PTT combined with a STING agonist (c-di-GMP) as an immune adjuvant can amplify the aroused antitumour immunity

Article Snippet: Evaluation of synergistic therapeutic effect of PTT combined with a STING agonist For further evaluation of the therapeutic effect of PTT combined with STING agonist c-di-GMP (HY-110382, MedChemExpress, NJ, USA), C57BL/6 N mice were injected subcutaneously with 5 × 106 Pan02-luc cells per mouse.

Techniques: Adjuvant

Journal: Journal of translational medicine

Article Title: Photothermal therapy combined with a STING agonist induces pyroptosis, and gasdermin D could be a new biomarker for guiding the treatment of pancreatic cancer.

doi: 10.1186/s12967-025-06247-2

Figure Lengend Snippet: Fig. 3 Pyroptosis induced by PTT combined with STING agonist and evaluation of anti-tumour effects of different therapies in Pan02-luc tumour-bearing mice. (a) Pan02-luc cells were divided into the Blank, PTT, c-di-GMP and c-di-GMP + PTT groups. Total proteins were extracted and subjected to western blotting analysis. (b) Morphological images of Pan02-luc after different treatments under a bright field microscope. Scale bar: 100 μm. (c) Schematic dia gram of the pancreatic tumour model and administration method. (d) The tumour volume during a 14-day observation post treatment (n = 5). (e) Survival analysis of mice during a 90-day observation post treatment (n = 5). (f) The body weight of mice during a 14-day observation post treatment (n = 5). (g) H&E staining of the major organs from the c-di-GMP + PTT treated mice. Scale bar: 100 μm. For analysis of tumour volume and body weight at 14 days post treatment, data were represented as mean ± SD if they accord with normal distribution. The comparison among different groups was carried out using one-way ANOVA followed by post hoc test. For survival analysis, Kaplan-Meier survival curves were conducted and the comparison between different groups was performed by Mantel-Cox test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Article Snippet: Evaluation of synergistic therapeutic effect of PTT combined with a STING agonist For further evaluation of the therapeutic effect of PTT combined with STING agonist c-di-GMP (HY-110382, MedChemExpress, NJ, USA), C57BL/6 N mice were injected subcutaneously with 5 × 106 Pan02-luc cells per mouse.

Techniques: Western Blot, Microscopy, Staining, Comparison

Journal: mBio

Article Title: Global Transcriptional Repression of Diguanylate Cyclases by MucR1 Is Essential for Sinorhizobium -Soybean Symbiosis

doi: 10.1128/mBio.01192-21

Figure Lengend Snippet: Overview of c-di-GMP signaling in Sinorhizobium fredii SF45436. (A) Synthesis of c-di-GMP by diguanylate cyclase (DGC) harboring the GGDEF domain and degradation of c-di-GMP by phosphodiesterase (PDE) containing the EAL domain. X indicates various sensory domains. The number of proteins, with each corresponding domain organization form, encoded by the SF45436 genome is shown (No.). Among them, the number of proteins with a conserved active motif [D X(7 aa) N X(8 aa) D X(21 aa) R X G /S/A GD /E EF] is indicated (Act). (B) The number of genes encoding proteins with putative activity of DGC or PDE, DGC-PDE bifunctional proteins, or PilZ-like proteins in Sinorhizobium sibling species. The point in orange corresponds to the value of SF45436. The other test Sinorhizobium strains include S. fredii SF25509, S. fredii SF83666, S. sojae SJ05684, and Sinorhizobium sp. strain SS05631. Error bars represent SD.

Article Snippet: The resultant 600-μl extraction solution was analyzed by HPLC-MS/MS with c-di-GMP (MedChemExpress, USA) as an internal standard.

Techniques: Activity Assay

Journal: mBio

Article Title: Global Transcriptional Repression of Diguanylate Cyclases by MucR1 Is Essential for Sinorhizobium -Soybean Symbiosis

doi: 10.1128/mBio.01192-21

Figure Lengend Snippet: Characterization of the GGDEF-containing proteins from S. fredii SF45436. (A) A sequence alignment of the GGDEF domain. A reported conserved motif essential for functional DGCs is shown. Gray background indicates other conserved amino acids in this sequence alignment (present in more than 80% sequences). Residues in black background form the reported allosteric I site involved in product inhibition of c-di-GMP synthesis. (B) Congo red binding ability of E. coli strains harboring various GGDEF-containing proteins from SF45436. Western blotting with anti-His monoclonal antibody shows the induced expression of corresponding proteins by IPTG. (+) and (−) indicate proteins with/without a conserved active motif [D X(7 aa) N X(8 aa) D X(21 aa) R X G /S/A GD /E EF]. YdeH is a known functional DGC from E. coli . For proteins not expressed in BL21(DE3) harboring pET28a(+) derivatives, the Rosetta strains (#) containing the same expressing plasmids or BL21(DE3) carrying pET30a derivatives ($) were tested. (C and D) HPLC-MS determination of c-di-GMP (C) or pGpG (D) content in 1 μM purified protein prepared from E. coli strains harboring corresponding vectors derived from pET30a or pET28a(+). Error bars represent standard deviations. Different letters above error bars indicate significant difference between means based on three independent experiments (ANOVA followed by Bonferroni’s multiple-comparison test, α = 0.05). (E) HPLC-MS determination of c-di-GMP content in E. coli strains expressing the indicated proteins relative to that of strains carrying the empty vector (*, P < 0.05; **, P < 0.01; ***, P < 0.001; t test). Error bars represent SD. Protein purification for these five proteins was not successful under test conditions, and in vitro enzymatic assay was not done.

Article Snippet: The resultant 600-μl extraction solution was analyzed by HPLC-MS/MS with c-di-GMP (MedChemExpress, USA) as an internal standard.

Techniques: Sequencing, Functional Assay, Inhibition, Binding Assay, Western Blot, Expressing, Purification, Derivative Assay, Comparison, Plasmid Preparation, Protein Purification, In Vitro, Enzymatic Assay

Journal: mBio

Article Title: Global Transcriptional Repression of Diguanylate Cyclases by MucR1 Is Essential for Sinorhizobium -Soybean Symbiosis

doi: 10.1128/mBio.01192-21

Figure Lengend Snippet: Symbiotic defects mediated by c-di-GMP overload in soybean nodules. (A) Chlorophyll content. (B) Nodule number per plant. Test strains include deletion (Δ) or complementary (.c) strains for DGC genes ( SFc17580 and SFb52570 ), the deletion mutant of PDE gene ( SFc33230 ), and derivatives carrying PDE ( SFc33230 EAL ) and DGC ( SFc17580 ) driven by the nifH ( SFa46030 ) promoter P nifH . The Δ mucR1 mutant was used for comparison. Error bars represent SEM. Different letters indicate significant differences between means based on more than 12 plants (ANOVA followed by Turkey’s multiple-comparison test; α = 0.05). (C) Pictures indicating yellow leaves and the increased nodule number of soybean plants inoculated with the P nifH -DGC and the Δ mucR1 mutant. (D) c-di-GMP levels in nodules (three biological replicates; error bars represent SD). (E) Transcriptional changes of SFc17580 and SFc33230 EAL in test strains (three biological replicates, ANOVA followed by Dunnett’s multiple comparisons; ***, P < 0.001; ****, P < 0.0001; error bars represent SD).

Article Snippet: The resultant 600-μl extraction solution was analyzed by HPLC-MS/MS with c-di-GMP (MedChemExpress, USA) as an internal standard.

Techniques: Mutagenesis, Comparison

Journal: mBio

Article Title: Global Transcriptional Repression of Diguanylate Cyclases by MucR1 Is Essential for Sinorhizobium -Soybean Symbiosis

doi: 10.1128/mBio.01192-21

Figure Lengend Snippet: Transcriptional changes of c-di-GMP responsive genes of the Δ mucR mutant and the P nifH -DGC and P nifH -PDE strains in soybean nodules. An iron-responsive regulator gene, rirA , is used as a control. Significant differences compared to the wild-type SF45436 are indicated (three biological replicates, ANOVA followed by Dunnett’s multiple comparisons; **, P < 0.01; ***, P < 0.001).

Article Snippet: The resultant 600-μl extraction solution was analyzed by HPLC-MS/MS with c-di-GMP (MedChemExpress, USA) as an internal standard.

Techniques: Mutagenesis, Control

Journal: mBio

Article Title: Global Transcriptional Repression of Diguanylate Cyclases by MucR1 Is Essential for Sinorhizobium -Soybean Symbiosis

doi: 10.1128/mBio.01192-21

Figure Lengend Snippet: Global repression of functional DGC genes located on chromosome and chromid by MucR in efficient soybean nodules. Genes encoding functional and nonfunctional DGCs are in red and blue, respectively. Artificial accumulation of c-di-GMP in nodules by using PnifH-c17580 leads to upregulation of various c-di-GMP responsive processes and impairs symbiotic performance. Other regulators can be involved in the coordinated regulation of these DGC genes under different conditions but have not been identified yet. Circular replicons (chromosome and chromid) are linearized here for presentation purpose.

Article Snippet: The resultant 600-μl extraction solution was analyzed by HPLC-MS/MS with c-di-GMP (MedChemExpress, USA) as an internal standard.

Techniques: Functional Assay