2.4. Chemotaxis and transwell cell migration assay

In the transwell top chamber (10 μg/mL; Costar, Cambridge, MA, USA) were added cells RPMI8226 and U266 (4.0 to 105 cells). The lowness of the chamber was supplemented by CXCL12 (250 ng/mL; R&D systems) or a 600 μL migration buffer alone. After three hours of incubation with a temperature of 37°, input cells and cells were analyzed using a FACSCalibur (BD-PharMingen) flow cytometer (Badr et al., 2011a, Badr et al., 2011b) and counted by flow cytometry. The data is shown as the percentage of CXCL12-induced migration input cells.