YDJC Induces Epithelial-Mesenchymal Transition via Escaping from Interaction with CDC16 through Ubiquitination of PP2A
Figure 4
CDC16 regulates EMT. (a) Effect of CDC16 overexpression on SPC-induced EMT. CDC16 overexpression was done by transfecting A549 cells with the plasmid containing CDC16 and the empty control vector (4 μg) followed by subsequent treatment with SPC (5 μM) for 48 h. (b) Confocal microscopic analysis of the effect of CDC16 overexpression on SPC-induced EMT. (c) Effect of CDC16 siRNA (siCDC16) on SPC-induced EMT. Gene silencing of CDC16 was accomplished by transfecting A549, H838, and H23 cells with the CDC16 siRNA and control siRNA and stimulation with or without 5 μM SPC for 48 h, respectively. (d) Confocal microscopic analysis of the effect of CDC16 siRNA (siCDC16) on SPC-induced EMT. (e) Effects of CDC16 siRNA and CDC16 overexpression on migration and invasion of A549 lung cancer cells. After siRNA of CDC16 and subsequent treatment with SPC, the A549, H838, and H1299 cells were added to the upper chamber of a Transwell insert to conduct the migration and invasion test. (f) Effects of CDC16 siRNA on changes of EMT markers in lung cancer cells. (g) Confocal microscopic analysis of the effects of CDC16 siRNA on changes of EMT markers in lung cancer cells. In (a), (c), and (f), cell lysates were analyzed by Western blotting.