tak875  (Cayman Chemical)

Journal: Diabetes & Metabolism Journal

Article Title: GPR40 Agonism Modulates Inflammatory Reactions in Vascular Endothelial Cells

doi: 10.4093/dmj.2021.0092

Figure Lengend Snippet: G protein-coupled receptor 40 (GPR40)-specific agonist can modulate nuclear factor-kappa B (NF-κB) activation under lipopolysaccharide (LPS) treatment. Cells were pretreated with LY2922470 (25 or 50 μM), TAK875 (25 or 50 μM), or LY2922470 plus GW1100 (10 μM) for 24 hours and then stimulated with LPS (200 ng/mL) plus the indicated reagents for 0.5 hour. (A, B) Phosphorylated NF-κB level was determined by Western blot. (C, D) Intracellular location of NF-κB was visualized through immunofluorescence microscopy analysis (×200). White arrows indicate that NF-κB was translocated from cytosol into the nucleus (red fluorescence, NF-κB; blue fluorescence, nucleus). (E, F) Western blot shows the level of nuclear NF-κB. Lamin B1 was used as the loading control for nuclear protein. The mean±standard deviation was obtained from three separate experiments. NE, nuclear extract. a P <0.05 vs. vehicle group, b P <0.05 vs. LPS group, c P <0.05 vs. LPS plus LY2922470 group, as analyzed with analysis of variance (ANOVA) followed by an un-paired Student’s t -test.

Article Snippet: The GPR40 agonists LY29-22470 and TAK875 and the GPR40 antagonist GW1100 (Cayman Chemical, Ann Arbor, MI, USA) were dissolved in dimethylsulfoxide (DMSO, Sigma Aldrich, St. Louis, MO, USA), and lipopolysaccharide (LPS, Sigma Aldrich) was diluted in phosphate-buffered saline.

Techniques: Activation Assay, Western Blot, Immunofluorescence, Microscopy, Fluorescence, Standard Deviation

Journal: Diabetes & Metabolism Journal

Article Title: GPR40 Agonism Modulates Inflammatory Reactions in Vascular Endothelial Cells

doi: 10.4093/dmj.2021.0092

Figure Lengend Snippet: G protein-coupled receptor 40 (GPR40)-specific agonist can regulate adhesion molecule expression under lipopolysaccharide (LPS) treatment. Human umbilical vein endothelial cells (HUVECs) were pretreated with LY2922470 (25 or 50 μM), TAK875 (25 or 50 μM), or LY2922470 plus GW1100 (10 μM) for 24 hours and then stimulated with LPS (200 ng/mL) plus the indicated reagents for 6 hours. (A, B) Vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) levels were determined by Western blot. (C, D) The stimulated HUVECs were co-cultured with green fluorescence-labeled THP-1 cells for 0.5 hour, and then THP-1 cells attached to HUVECs were analyzed using a fluorescence microscope (×200) and a spectrofluorometer. The mean±standard deviation was obtained from three separate experiments. a P <0.05 vs. vehicle group, b P <0.05 vs. LPS group, c P <0.05 vs. LPS plus LY2922470 group, as analyzed with analysis of variance (ANOVA) followed by an un-paired Student’s t -test.

Article Snippet: The GPR40 agonists LY29-22470 and TAK875 and the GPR40 antagonist GW1100 (Cayman Chemical, Ann Arbor, MI, USA) were dissolved in dimethylsulfoxide (DMSO, Sigma Aldrich, St. Louis, MO, USA), and lipopolysaccharide (LPS, Sigma Aldrich) was diluted in phosphate-buffered saline.

Techniques: Expressing, Western Blot, Cell Culture, Fluorescence, Labeling, Microscopy, Standard Deviation