The suppressive effect of IPA-3 on migration of H2M cells.
(A) Paxillin protein expression was detected by immunofluorescence analysis under IPA-3 treatment. H2M and H2P (upper panel) cells were serum-starved overnight and treated with either DMSO control or IPA-3 (20 µM) for 15 minutes, followed by FBS replenishment for 10 minutes. Immunofluoresence signals of phalloidin (Red), paxillin (Green) and DAPI (Blue) represent stress fiber, focal adhesion and nucleus, respectively (magnification 40X). The number of focal adhesion (paxillin) were counted in H2M (lower, left panel) and H2P (lower, right panel), and represented in the bar chart. Error bars, mean ± SD of triplicate samples. *P<0.01 (t-test) compared with DMSO control. (B) Western blotting analysis on the phosphorylation levels of PAK1 and paxillin. Serum-starved cells were treated with various concentrations of IPA-3 as indicated for 15 minutes, followed by FBS replenishment for 10 minutes. (C) Representative images of Transwell migration assay of H2M cells. Cells were treated with either DMSO or 10 µM IPA-3, and were allowed to migrate for 24 hours. Images show the cells having migrated to the lower chamber (upper panel). The number of migrated cells were counted and represented in the bar chart (lower panel). Error bars, mean ± SD of triplicate samples. *P<0.01 (t-test) compared with DMSO control.