Fig. 6

AC-93253 iodide inhibited Src and Src-related gene transcription and induced protein degradation. a AC-93253 iodide induced increases in protein degradation. Western blot analyses of Src, EGFR, STAT3, and FAK expression levels in PC9 cells treated with or without the protein synthesis inhibitor CHX and/or AC-93253 iodide for 4, 8, and 12 h. b AC-93253 iodide induced increases in ubiquitination, as determined by western blotting. The proteasome inhibitor MG132 and/or AC-93253 iodide were administered to PC9 cells for 72 h. GAPDH served as a loading control. c Repressive effects of AC-93253 iodide on Src, EGFR, STAT3, and FAK transcription as determined by real-time RT-PCR in PC9 cells. Relative gene expression levels were calculated using the comparative CT method (2^–ΔΔCT). TBP: internal control. Each experiment was performed independently and in triplicate. *P < 0.05 compared with control (0 nM: 0.1% DMSO). d Enhanced ubiquitination of Src-related proteins induced by AC-93253 iodide. PC9 cell lysates with or without AC-93253 treatment were immunoprecipitated by the indicated antibodies and then analyzed by western blotting with anti-ubiquitin. GAPDH served as the internal control. Con represents 0 nM (0.1% DMSO)