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Image Search Results

Journal: Frontiers in Immunology
Article Title: Cholinergic Signaling Attenuates Pro-Inflammatory Interleukin-8 Response in Colonic Epithelial Cells
doi: 10.3389/fimmu.2021.781147
Figure Lengend Snippet: Cholinergic stimulation attenuates LPS-induced IL-8 response in SW480 cells via a7nAChR. Expression of ACHRs in SW480 cells (A) . SW480 cells (5 x 10 4 /200µl) were stimulated with LPS (100ng/ml) in the presence of different agonist and antagonists. IL-8 was measured in cell-free supernatants 20h after LPS stimulation. Cells were stimulated with LPS 20min prior to acetylcholine (ACh, 10µM), nicotine (10µM), and muscarine (10µM) stimulation (B) . Cells were pretreated (20min) with the non-selective nAChR antagonist Mecamylamine (Mec, 100µM) followed by treatment with ACh (10µM), 20min prior to LPS stimulation (C) . Cells were pretreated (20min) with the selective a7nAChR antagonist alpha-Bungarotoxin (a-BTX, 100µM) followed by treatment with GTS-21 (100µM), 20min prior to LPS stimulation (D) . Cells were pretreated (20min) with GTS-21 (100µM) followed by stimulation with LPS. Phospho (p)-NFĸB-p65 was measured 4h after LPS stimulation (E) . Cells were pretreated (20min) with DMSO (0.01%, control) or tyrphostin AG490 (1µM) or wortmannin (1µM) followed by stimulation with LPS (100ng/ml), 20min prior to treatment with a7nAChR agonist GTS-21 (20min) (F) . Cell viability was assessed by colorimetric MTS assay (G) . Each point in the scatter plots with bar represent triplicates or quadruplicates ± SEM. Data are representative of three independent experiments. Significance was determined using one-way ANOVA multiple comparison analysis (A–D) and paired t test (E, F) with *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.005. ns, not significant. n.d., not detectable.
Article Snippet: The following agonists/antagonists were used: acetylcholine chloride (10µM, AChR agonist, Merck, Darmstadt, Germany), nicotine (10µM, nAChR agonist, Merck, Darmstadt, Germany), muscarine chloride hydrate (10µM, mAChR agonist, Merck, Darmstadt, Germany), GTS-21 (100µM, a7nAChR agonist, Merck, Darmstadt, Germany), mecamylamine HCL (100µM, nAChR antagonist, Merck, Darmstadt, Germany),
Techniques: Expressing, MTS Assay