Author: Wu, Beibei; Wang, Liyin; Jiang, Lili; Dong, Lili; Xu, Fengli; Lu, Yili; Jin, Jiahui; Wang, Zhanyue; Liang, Guang; Shan, Xiaoou
Title: n-butanol extract from Folium isatidis inhibits the lipopolysaccharide-induced downregulation of CXCR1 and CXCR2 on human neutrophils Document date: 2017_10_25
ID: w85t4zz6_8
Snippet: Chemotaxis assays. The isolated neutrophils were pretreated as described above for 2 h and then stimulated with LPS (0.5 µg/ml) for 4 h. The cells were then washed twice with PBS, and resuspended in RPMI 1640 at 1x10 6 cells/ml. The chemotaxis assays were performed using Transwell inserts (24 wells, 5 µm pore size; Corning, Inc., Corning, NY, USA), where 10,000 cells in 100 µl were loaded in the upper inserts and 600 µl of IL-8 (100 ng/ml; eB.....
Document: Chemotaxis assays. The isolated neutrophils were pretreated as described above for 2 h and then stimulated with LPS (0.5 µg/ml) for 4 h. The cells were then washed twice with PBS, and resuspended in RPMI 1640 at 1x10 6 cells/ml. The chemotaxis assays were performed using Transwell inserts (24 wells, 5 µm pore size; Corning, Inc., Corning, NY, USA), where 10,000 cells in 100 µl were loaded in the upper inserts and 600 µl of IL-8 (100 ng/ml; eBioscience; Thermo Fisher Scientific, Inc.) containing RPMI 1640 was added to the lower wells. Following co-incubation for 2 h at 37˚C and a humidified 5% CO 2 atmosphere, the inserts were removed, and the migrated neutrophils in the lower wells were collected and quantified according to myeloperoxidase (MPO) activity using a myeloperoxidase assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
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