FIGURE 2:
Analysis of PrPc, γ-catenin, β-catenin, and TCF7L2 (TCF4) levels in proliferating Caco-2/TC7, HIEC-6 and SW480 cells. (A) Western blot analysis of total protein extracts from Caco-2/TC7, HIEC-6, and SW480 cells (2 d after plating). A 30-μg amount of protein was analyzed for PrPc, γ-catenin, β-catenin, and TCF7L2 levels in each cell line in triplicate, with actin as loading control. Bar graphs show the ratio of each protein to actin after densitometric analyses (mean ± SEM; *p < 0.05, **p < 0.01 ***p < 0.001 vs. Caco-2/TC7; $p < 0.05, $$$p < 0.001 vs. SW480). (B) Western blot analysis of nuclear (Nu) and cytoplasmic (Cyt) extracts of the three cell lines. A 30-μg amount of protein was analyzed for PrPc, γ-catenin, and β-catenin levels in each fraction and each cell line. Purity of nuclear and cytoplasmic fractions was checked by SP1 and LDH analyses; *nonspecific band revealed by the anti-SP1 antibody.