24-Well Cell Invasion Assays, Basement Membrane

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 3 × 10^5 cells	Incubate for 24hours at 37ºC in 5% CO2 atmosphere.	Measure the OD 560nm in a plate reader

Upstream tips
Seed 3 × 10^5 cells
Protocol tips
Incubate for 24hours at 37ºC in 5% CO2 atmosphere.
Downstream tips
Measure the OD 560nm in a plate reader

Publication protocol

Cell invasion was evaluated with the CytoSelect 24-Well Cell Invasion Assay kit (Cell Biolabs, San Diego, CA). After siRNA knockdown, the cells were cultured in 10−9 M E2-containing medium for 36 h and starved in serum free medium for overnight. Then, 3 × 105 cells in serum-free medium were seeded on a basement membrane-coated insert with 8 μM pores, and 500 μl of medium containing 10% of charcoal-stripped FBS and E2 (10−9 M) were added to the lower chamber. After a 24-h incubation, cells that had penetrated the insert pore were extracted and absorbance at OD560nm was measured.

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