Figure 3.

ZY‐444 directly bound to PC and decreased the enzymatic activity of PC. A) The chemical structure of biotin‐conjugated ZY‐444 (Biotin‐ZY‐444). B) The comparative effects of ZY‐444 and biotin‐ZY‐444 on cell migration. C) MDA‐MB‐231 cell lysates were incubated with biotin or biotin‐ZY‐444 overnight. Streptavidin‐conjugated agarose beads were used to precipitate their binding proteins, which were separated by gel electrophoresis and visualized by silver staining. The arrowhead indicates a specific band in the gel, which was identified as PC through mass spectrum analysis. D) The biotin terminal end of biotin‐ZY‐444 was immobilized on the sensor chip. It interacted with the indicated concentrations of purified PC. Strong drug–protein interaction resulted in a series of binding response curves. E,F) PC expression after ZY‐444 exposure for 24 h at E) the mRNA and F) protein levels in MDA‐MB‐231 cells. G) The effects of Octyl gallate and ZY‐444 on PC enzymatic activity. H) A microarray gene expression profiling analysis was performed using MDA‐MB‐231 cells treated with vehicle versus ZY‐444. PANTHER classification analysis provided the percentages of gene hits against total biological hits by ZY‐444. Data shown are mean ± s.d. p < 0.001, ***.