Effect of berberine and As
on the translocation of PKCα and PKCε. C6 glioma cells were incubated for 12 and 24 h with 10 μM berberine (Ber) or 5 μM As2O3 (As), and with 10 μM berberine and 5 μM As2O3 (A+Ber) (A, B). Cytosolic (B) and membrane fractions (A) were evaluated for the presence of PKCα and PKCε by Western blotting. Control cells were not treated in the presence of 10% Fetal Calf Serum (FCS) (N10). Cells that were not exposed to agents were used as negative control in the presence of 1% FCS (N1). The PKC levels in the respective fractions were quantified and normalized taking the β-actin value as a loading control as presented in the graph. Each value is the relative ratio of PKCs membrane to cytosol fraction (presumably, the ratio of untreated control is 1). In addition, the nuclear protein fractions were evaluated for the presence of myc and c-jun, two transcription factors that act downstream of PKC, by Western blotting (C). The quantitative data were presented as three repeats from one independent experiment and indications are in panel A. The intensity of the protein bands was quantified and the level relative to the N10 control band was presented (fold).