Cal inhibits invasion by suppressing PKC-α. (A and B) Cells were pretreated with AEB071 (0.1 µM) for 30 min and then incubated in the presence or absence of Cal (30 µM) for 24 h. The A549 cells were then subjected to western blotting to analyze the protein levels of PKC-α and (C and D) E-Cad, integrin β1, MMP-2 and MMP-9. (E) Cells were pretreated with AEB071 (0.1 µM) for 30 min and then incubated in the presence or absence of Cal (30 µM) for 24 h. Cellular invasiveness was measured using a Transwell invasion assay and an IX73 microscope following staining with crystal violet (magnification, ×100). (F) Invasion rate is expressed as a percentage of the TPA-induced group. Values are presented as the mean ± standard deviation of three independent experiments, performed in triplicate. *P≤0.05 and **P≤0.01, vs. TPA-induced group. #P≤0.05 and ##P≤0.01, vs. Cal+ group. Cal, calycosin; TPA, phorbol-12-myristate-13-acetate; PCK, protein kinase C; E-Cad, E-cadherin; MMP, matrix metalloproteinase.