Matrigel invasion inserts

Cell migration of MDA-MB231 and MCF-7 cells was detected with a 24-well Matrigel™ Invasion inserts (8 μm pore size; BD Biosciences, Franklin Lakes, NJ, USA) as described elsewhere [32 (link)]. Serum-free medium containing different concentrations of T33 and 2 × 10⁴ cells were loaded into the upper chamber whereas the 10% fetal bovine serum (FBS)-containing medium was loaded into the lower chamber. After 24 h incubation, 10% neutral-buffered formalin was used to fix the migrated cells on the lower surface of the membrane and the migrated cells were then stained with 0.5% crystal violet for 15 min. The stained cells were then counted in five randomly selected microscopic fields at 200× magnification per filter.

A cell invasion assay was conducted in 24‐well Matrigel™ Invasion inserts (8 μm pore size; BD Biosciences, Franklin Lakes, NJ, USA) to measure the effects of escitalopram oxalate. Briefly, 2 × 10⁴ cells in serum‐free DMEM that contained various concentrations of escitalopram oxalate were loaded into the upper chamber and DMEM that contained 10% foetal bovine serum was added to the lower chamber. After 2 days of incubation, the cells on the upper surface of the membranes were removed using a cotton swab. Migrated cells on the lower surface of the membrane were fixed with 10% neutral‐buffered formalin and stained with 0.5% crystal violet for 15 min. Three independent assessors were then performed by counting the cells in five randomly selected microscopic fields at 200× magnification per filter.