|Incubate for 4-6 hours in a cell culture incubator
The experiment was carried out using the CytoSelect cell migration assay kit (Cell Biolabs, Inc., San Diego, CA) containing polycarbonate membrane inserts (8 μm pore size) in a 24-well plate. In brief, appropriate amount of cells were suspended in serum-free medium and added to the insert. The lower well was loaded with routine medium (with 10% FBS). After an incubation of 4-6 hours at 37°C in a humidified incubator with 5% CO2, non-migratory cells on the upper surface of the membrane were gently removed by wiping with a cotton swab. The cells that migrated through the pores and to the underside of the membrane were fixed, stained and photographed under a microscope. The number of cells was counted and averaged using three randomly selected areas per membrane. Doxycycline (0.8 µg/mL) was added to medium/cells over the time course of the desired experimental groups. Full paper
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