Inhibitory effect of AT13387 on C666-1 cell migration. (A) Trans-well migration assay. The C666-1 cells treated with AT13387 for 72 hrs were harvested and viable cells were seeded on the upper chamber of transwell for migration assay. Upper panel: images of cells migrated through the membrane of migration chamber stained with DAPI. Scale bar = 100 μm. Lower panel: quantification of migrated cells. At least 100 cells were counted from different microscopic fields. Results were expressed as the mean ± S.D. of three separate trials; *p < 0.05. (B) Western-blotting analysis of C666-1 treated with AT13387 for 72 hrs and 96 hrs showed decreased expression of cell migration regulator HDAC6 and increased acetylated form of the α-tubulin. Value represented the arbitrary unit of band intensity after normalization with β-actin.