Journal: The FASEB Journal

Article Title: A non‐polar fraction of Saponaria officinalis L. acted as a TLR4/MD2 complex antagonist and inhibited TLR4/MyD88 signaling in vitro and in vivo

doi: 10.1096/fj.202200018rr

Figure Lengend Snippet: FIGURE 7 fr.1 and saponarin inhibited the LPS-mediated interaction between TLR4 and MyD88 in THP-1 macrophages. Relative TLR4/MyD88 interactions in LPS-induced macrophages were determined by western blot of immunoprecipitation samples. (A) Western blot images of TLR4-co-immunoprecipitated (Co-IP) protein samples obtained from fr.1 or saponarin treated THP-1 macrophages. Co-IP proteins were detected using TLR4 or MyD88 antibodies. (B) Relative ratios of TLR4 interacting with MyD88 adaptor protein. Band intensities were measured by densitometry and normalized versus TLR4 band intensities. Results are presented as the means ± SDs of three different experiments. #p < .05 versus non-treated control macrophage group, and *p < .05 versus LPS-treated inflammatory macrophage group.

Article Snippet: Primary antibodies for iNOS (Inducible nitric oxide synthase), COX- 2 (Cyclooxygenase- 2), TLR4 (Toll- like receptor 4), MyD88 (Myeloid differentiation primary response 88), lamin B, and β- actin, and horseradish peroxidase (HRP)- conjugated secondary antibodies were purchased from Santa Cruz (Dallas, TX, USA).

Techniques: Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Control

Journal: The FASEB Journal

Article Title: A non‐polar fraction of Saponaria officinalis L. acted as a TLR4/MD2 complex antagonist and inhibited TLR4/MyD88 signaling in vitro and in vivo

doi: 10.1096/fj.202200018rr

Figure Lengend Snippet: FIGURE 10 Molecular docking studies of the interaction between saponarin and TLR4/MD2 complex. (A) Depiction of ligand clusters of saponarin or dexamethasone binding to TLR4/MD2 complex as deduced by protein-ligand docking simulation in Swissdock. (B) Estimated binding energy and pose of the best MD2-ligand binding simulation model. UCSF chimera depicted MD2 protein as a ribbon and provided a surface image. (C) Sidechains of MD2 protein proximal to ligands (within 5 Å) were labeled and visualized by UCSF chimera. (D) 2D image of non-ligand residues and its corresponding atoms of ligand visualized by Ligplot plus.

Article Snippet: Primary antibodies for iNOS (Inducible nitric oxide synthase), COX- 2 (Cyclooxygenase- 2), TLR4 (Toll- like receptor 4), MyD88 (Myeloid differentiation primary response 88), lamin B, and β- actin, and horseradish peroxidase (HRP)- conjugated secondary antibodies were purchased from Santa Cruz (Dallas, TX, USA).

Techniques: Binding Assay, Ligand Binding Assay, Labeling