Bosutinib inhibit cell migration and invasion is ACK1 dependent and SRC- independent. NCI-H1792 were transfected with siRNA (control, ACK1 or SRC) using Oligofectamine for 72 h. The cells were harvested at 72 h. (A) Quantitative real-time PCR analysis was performed using 20 ng of cDNA and respective primers set for ack1 and src and normalized to gapdh. The serum-starved cells were trypsinized and seeded in upper chamber of the Transwell (8 mm pore, B) or Matrigel™ (C), in the presence of DMSO or bosutinib at various concentrations (0.5 and 1 μM). Medium containing 10% FBS and DMSO or bosutinib (0.5 and 1 μM) was used as chemoattractant in the lower chamber. The cells were fixed and stained with 0.5% crystal violet blue after 6 h (migration, B) and 24 h (invasion, C). Cells that migrated or invaded across the filter were counted. Experiments were carried out in duplicates with five random fields counted.