Figure 4

Effects of MRBE on ATF3 and cyclin D1 expression in mRNA and protein level in SW480 cells. (A, B) SW480 cells were treated with MRBE at the indicated concentrations for 24 h. Cell lysates were subjected to SDS–PAGE and the Western blot was performed using antibodies against cyclin D1, ATF3 and actin. For RT-PCR analysis of ATF3 and cyclin D1 gene expression (C), total RNA was prepared after MRBE treatment for 24 h. (D) SW480 cells were treated with MRBE (25 μg/ml) for the indicated times. Cell lysates were subjected to SDS–PAGE and the Western blot was performed using antibodies against cyclin D1, ATF3 and actin. Actin and GAPDH were used as internal control for Western blot and RT-PCR, respectively. DMSO was used as a vehicle.