The conventional invasion assay was performed using a BD BioCoat Growth Factor Reduced Matrigel Invasion Chamber (354483, BD Biosciences, NJ, USA) as described previously42 (link). In brief, cells, which were plated on the upper insert at 1 × 105/well with or without nanoparticles, were incubated for 24 h. The invaded cells on the under surface of the membrane were fixed after removing the cells on the upper surface of the membrane in each insert. The number of invaded cells, which were stained with 0.05% Toluidine Blue Solution (206-14555, Wako), was manually counted by microscopy.
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