Effect of AR inhibition on hypoxia-mediated and/or FGF- or serum-mediated PI3K/AKT, GSK3β, and Snail activities and invasion of HT29 cells. Growth-arrested HT29 cells were preincubated with fidarestat (Fid) or carrier for 24 h, followed by stimulation under hypoxia and/or FGF for another 24 h. A, Western blot analysis performed using antibodies against phospho-p85α, phospho-AKT, phospho-GSK3β, phospho-Snail, and GAPDH. Fold values of normalized intensity measured by densitometric scanning. B, PI3K activity was measured after 6 h of hypoxia and/or FGF or serum treatment. C, growth-arrested HT29 cells were plated at 20,000 cells/well in a 96-well plate containing Matrigel and treated with hypoxia and/or FGF (10 ng/ml) or serum with or without fidarestat (5 μm). After an overnight incubation, the invasion of cells toward the bottom side of the well was measured in situ using calcein-AM florescent dye at 480/520 nm. Bars represent mean ± S.E. (n = 4). *, p < 0.001 compared with control (Con) and #, p < 0.001 compared with cells treated with hypoxia and/or FGF or serum.