Transwells 8 μm pore pet membrane

Cell invasion assays were performed using Matrigel (BD Transduction, San Jose, CA, USA)-coated Transwells (8 μm pore PET membrane, Millipore, Billerica, MA, USA) as previously described (19). Briefly, 2.5 μM Sorafenib or Regorafenib treated cells were suspended in low serum medium. Medium containing different hPL or FBS concentrations was added to the bottom wells. After incubation of 24 h, the invading cells were fixed and stained. The images were acquired and analyzed counting the cells with Image J Software (National Institute of Health, USA). Values obtained were expressed as fold increase of invading cells, setting the cell counts of control cells as one. Results were representative of three independent experiments.

Cell invasion assays were performed using Matrigel (BD Transduction, San Jose, CA, USA)-coated Transwells (8 μm pore PET membrane, Millipore, Billerica, MA, USA) as previously described [31 (link)]. Briefly, Regorafenib 5 μM and/or GSK1838705A 1 μM treated cells were suspended in low serum medium. Medium containing IGF 40 ng/ml or FBS was added to the bottom wells. After incubation of 24 h, the invading cells were fixed and stained. The images were acquired and analyzed counting the cells with Image J Software (National Institute of Health, USA). Values obtained were expressed as percentage of invading cells, setting the cell counts of control cells as 100 %. Results were representative of three independent experiments.