MELK could regulate cell migration and invasion via the FAK/Paxillin pathway. A and B, Effects of MELK on FAK and paxillin phosphorylation levels. Cellular lysates were analyzed by immunoblotting. C and D, 3′UTR-siRNA of MELK partially reversed the up-regulation of pY397, pY576/577, and pY925 of FAK, and pY118 of paxillin caused by MELK overexpression. SGC7901/vector and SGC7901/MELK cells were transfected with 3′UTR-siRNA of MELK using Lipofectamine 2000; cells were collected and immunoblotting was performed 48 h later. E, Effect of FAK inhibitor (FAK-I) on SGC7901 cell migration and invasion. SGC7901, SGC7901/vector and SGC7901/MELK cells were treated with FAK inhibitor (10 μM); cell migration and invasion were measured after 2 h. F, Knockdown of MELK prevents gastrin-stimulated FAK and paxillin phosphorylation. NCI-N87/NC-shRNA and NCI-N87/MELK-shRNA cells were treated with gastrin (200nM) and cells were collected and analyzed by immunoblotting after 0, 15, 30, 45, 60 mins. G, Ratio of phosphorylated to total protein, respectively. (Pax: paxillin).