Figure 3

ABT-263 increases the mRNA stability of Mcl-1. (A) Schematic representation of Mcl-1 promoter region containing several transcription factor binding sites. Two fragments (−3009 to +251 and −607 to +251) were separately amplified from genomic DNA of HepG2 cells and inserted into pGL3-basic vector, and the resulting plasmids were named as pLucM1 and pLucM2 respectively. (B) After co-transfected with pLucM1 or pLucM2 and pCMV-β-gal plasmid for 12 h, the HCC cells were treated with 5 μM ABT-263 or vehicle DMSO for another 24 h. Then the luciferase assay was performed. Data were expressed as mean ± SD from three independent experiments. (C) HCC cells were treated with 10 μg/ml actinomycin D (Act D) in the presence or absence of 5 μM ABT-263 for indicated times, then the level of Mcl-1 mRNA was quantified by qPCR, taking β-actin as a control. Data were represented as mean ± SD from three independent experiments. (*: P < 0.05).