Figure 1
GCDA activates ERK1/2 and induces Mcl-1 phosphorylation in HepG2 cells. (A) Expression of Mcl-1 in various HCC cell lines and normal liver cell line L02 was analyzed by Western blot. (B) HepG2 cells were treated with increasing concentrations of GCDA for 30 min. Phosphorylation of ERK1/2 or total ERK1/2 was analyzed by Western blot using phospho-specific ERK or ERK1/2 antibodies, respectively. (C) HepG2 cells were metabolically labeled with 32P-orthophosphoric acid for 60 min and treated with GCDA (100 μM) in the absence or presence of PD98059 (10 μM) for 30 min. Mcl-1 was immunoprecipitated by using Mcl-1 antibody. Phosphorylation of Mcl-1 was determined by autoradiography. Western blot analysis was performed to confirm and quantify Mcl-1 protein. Ctrl, immunoprecipitation with normal rabbit IgG as a negative control.