Journal: Frontiers in Immunology

Article Title: Key Components of the Complement Lectin Pathway Are Not Only Required for the Development of Inflammatory Arthritis but Also Regulate the Transcription of Factor D

doi: 10.3389/fimmu.2020.00201

Figure Lengend Snippet: Microscale thermophoresis showing biophysical analysis of binding of human rMBL with human rTLR4/MD2. MST is based on the detection of a temperature-induced change in fluorescence of rTLR4/MD2 (target) as a function of the concentration of a non-fluorescent ligand (rMBL). By titrating MBL into the labeled TLR4 the K d (dissociation constant) was 9.07 × E −07 indicating strong binding.

Article Snippet: Recombinant Human TLR4 (R&D Systems) was re-suspended in a buffer containing 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , and 1 mM BME.

Techniques: Microscale Thermophoresis, Binding Assay, Fluorescence, Concentration Assay, Labeling

Journal: Frontiers in Immunology

Article Title: Key Components of the Complement Lectin Pathway Are Not Only Required for the Development of Inflammatory Arthritis but Also Regulate the Transcription of Factor D

doi: 10.3389/fimmu.2020.00201

Figure Lengend Snippet: Effect of human rMBL or human rTLR4 on FD expression on differentiated 3T3-L1 cells at 48 h. (A) rMBL increased FD expression in a dose-dependent manner. (B) rMBL also effected the TLR4 expression. (C) LPS also increased FD expression dose-dependent manner. (D) LPS also decreased TLR4 expression with increasing doses. Data are shown as Mean ± SEM of three replicative experiments. * p < 0.05 considered significant.

Article Snippet: Recombinant Human TLR4 (R&D Systems) was re-suspended in a buffer containing 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , and 1 mM BME.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Key Components of the Complement Lectin Pathway Are Not Only Required for the Development of Inflammatory Arthritis but Also Regulate the Transcription of Factor D

doi: 10.3389/fimmu.2020.00201

Figure Lengend Snippet: A hypothetical model in mouse showing how MBL or conjugates of MBL-MASP-1 or MBL-MASP-2 can regulate the transcription of FD to regulate the activation of the AP via TLR4 receptors. Circulating MBL alone or MBL-MASP-1 or MBL-MASP-2 complexes can directly interact with disguised TLR4 on adipocytes to activate the complement system via enhancing the expression of FD to regulate the AP pathway. (A) MBL or MBL-MASP-1 or MBL-MASP-2 conjugates under normal physiological conditions can bind to the TLR4 and regulate the expression of FD but MASP-2 might be dominant. (B) In contrast, under inflammatory conditions, MBL or MBL-MASP-1 or MBL-MASP-2 conjugates might be displaced by the LPS and modulate the transcription of FD through TLR4 receptors.

Article Snippet: Recombinant Human TLR4 (R&D Systems) was re-suspended in a buffer containing 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , and 1 mM BME.

Techniques: Activation Assay, Expressing

Journal: The Journal of biological chemistry

Article Title: DNA-mediated proteolysis by neutrophil elastase enhances binding activities of the HMGB1 protein.

doi: 10.1016/j.jbc.2022.102577

Figure Lengend Snippet: Figure 6. Potential roles of the DNA-mediated proteolytic processing of HMGB1 by neutrophil elastase in NETs. Due to the enhanced binding activities of the processed HMGB1 protein, this processing may promote (1) TLR4 signaling, (2) binding to biofilm DNA, and (3) DNA sensing by cGAS. Due to the loss of residues 177–215, the processing of HMGB1 may diminish (4) RAGE signaling and (5) nuclear localization. NET, neutrophil extracellular trap.

Article Snippet: Lyophilized TLR4 MD-2 complex was purchased from R&D Systems (catalog no.: #3146-TM-050).

Techniques: Binding Assay