Transwell insert polycarbonate membranes

Migration assays were performed using transwell insert polycarbonate membranes (8 Mm pore size, BD, Franklin Lakes, NJ, USA). To test cell migration, 1×10⁵ cells of SiHa or C33A treated with SFA gels, matrine and oxymatrine at different concentrations and their corresponding controls were added in upper chamber and 600 μL DMEM with 10% fetal bovine serum was added to the bottom at 37 ºC for 24 hours. Nonmigratory cells on the upper membrane surface were removed, and migratory cells attached to the bottom surface of the membrane were then fixed in 100% methanol for 30 minutes and stained with crystal violet for 10 minutes. All the tests were performed three times.

Chemotaxis assays were performed using Transwell insert polycarbonate membranes (8 μm pore size, BD, Franklin Lakes, NJ, USA). First, 1×10⁵ cells were seeded in the top compartment and 600 μL DMEM with 10% fetal bovine serum was added to the bottom at 37°C for 13 hours. Nonmigratory cells on the upper membrane surface were removed, and migratory cells attached to the bottom surface of the membrane were fixed with 10% formalin and stained with 0.1% crystal violet for 30 minutes at room temperature. Migrated cells in five random fields were counted using a light microscope at 200× magnification. All assays were performed in triplicate.