Chemotaxis assays were performed using a transwell chamber (Costar) as previously described (24 (link)). The numbers of cells that migrated to the lower well after 2 h or 3 h incubation were counted using a MACSQuant flow cytometer (Miltenyi Biotec). The percent migration was calculated by the numbers of cells of a given subset that migrated into the bottom chamber divided by the total number of cells of that subset in the starting cell suspension, and multiplying the results by 100. D-erythro-sphingosine 1-phosphate was purchased from Avanti Polar Lipids. CCL19 and CXCL12 were purchased (R&D Systems). Fatty acid free bovine serum albumin (FAF-BSA) was purchased (Sigma-Aldrich).
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Chemotaxis Assay Using Transwell
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Corresponding Organization :
Other organizations : National Institute of Allergy and Infectious Diseases, National Institutes of Health
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Variable analysis
independent variables
- D-erythro-sphingosine 1-phosphate
- CCL19
- CXCL12
- Percent migration of cells into the bottom chamber
- Transwell chamber (Costar)
- Incubation time (2 h or 3 h)
- MACSQuant flow cytometer (Miltenyi Biotec)
- Fatty acid free bovine serum albumin (FAF-BSA)
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