To determine chemotaxis by S1P, CCL19, or CCL21 of Treg cells from WT and CCR7−/− mice, Cultrex 96 Well Cell Migration Assay Kit (Trevige, Inc., Gaithersburg, MD) was used in accordance with the manufacture's instructions.
Transmigration assay was performed using the QCM Chemotaxis migration assay kit (Millipore) according to the manufacturer's instruction.
A colorimetric cell migration assay kit (Cell Biolabs, San Diego, CA) consisting of 24-well plates that contain polycarbonate membrane filter inserts (8 μM pore size) was used according to the vendors's instructions.
Cell invasion was measured with a CytoSelect Cell Migration assay kit (Cell Biolabs Inc .. Cells were starved overnight in serum-free medium and resuspended at a concentration of 2.5 × 10 cells per ml in serum-free medium containing 0.1% BSA.
The ability of Rokumigan to promote wound healing was assessed using the Oris Pro Cell Migration Assay kit (Platypus Technologies, Madison, WI, USA) according to the manufacturer's protocol.
Cell invasion and migration assay kits were obtained by Chemicon, Millipore, Temecula, CA, USA.
In the enriched miRNAs, miR-221/222 is a well-studied miRNA which can significantly reduce tube formation and migration by directly targeting KIT (c-kit) [ 44, 45].
