*3.12. Chemotaxis Assay toward Eotaxin*

Eosinophils (1.3 × 106 cells/mL) were suspended in RPMI 1640 medium containing 0.1% BSA and were placed in the upper well (Transwell) of a 24-well chemotaxis chamber (KURABO Co., Osaka, Japan). Transwells with 5 ΐm pores were inserted into each well, and 4 × 105 cells in 300 ΐL of RPMI 1640 medium containing 0.1% BSA were added to the upper chamber. The lower chamber was set with 800 ΐL of RPMI 1640 medium containing 0.1% BSA and mouse eotaxin (20 ng/mL). Then, 3 or 1 ΐg of peridinin or fucoxanthin (*<sup>n</sup>* = 6) was added to each Transwell. Assay plates were incubated for 1 h at 37 °C in 5% CO2. Cells that migrated across the filter to the lower chamber were counted, and the results were expressed as the number of cells in a field of 660 ΐm × 840 ΐm. For each group, eosinophils in three fields for each well were counted, and results were reported as the mean of 18 fields (cell number ± SD). 
