Fig. 5

Effect of ethanol on CKII expression and/or activity in vitro and in vivo. a phosphorylated and unphopshorylated STARD10 in ethanol-treated MCF-7 cells protein extracts (100 mM for 48 h). STARD10 protein levels were analyzed by Western Blotting . The results are expressed as fold relative to control from 3 independent experiments and normalized with red pounceau. b, c CKII mRNA and protein levels were measured by RT-PCR and Western blotting, respectively in MCF-7 and SKBR-3 cell lines. CKII protein was normalized with β-actin expression. Three independent experiments were performed in triplicates. d The CKII enzymatic activity was evaluated as absorbance at 450 nm of cell lysate in modified kinase reaction buffer and normalized with standard in MCF-7 ans SKBR-3 cells treated with 100 mM ethanol for 48 h. Data are expressed as (mean ± SE) from 4 independent experiments performed in triplicates. *p < 0.01 vs. control. e Western blotting analysis of CKII in ethanol-fed MMTV-neu mice tumors. Densitometric ratios normalized to actin is shown in the right panel. Four mice per group were used. f CKII enzymatic activity assay in MMTV-neu was normalized with standard from 4 mice per group. *p < 0.005 vs. control tumor