Flotillin-2 knockdown increases raft association of E-cadherin and γ-catenin in MCF10A cells.

(A) Lipid rafts were isolated using detergent extraction from MCF10A control and flotillin-2 knockdown (sh-F2-A/B) cells after 10 days of confluent growth. Fractions 1–12 were stained with antibodies against E-cadherin, γ-catenin, flotillin-2, flotillin-1, GAPDH and transferrin receptor (TfnR). Lipid raft fractions were detected with an HRP-coupled cholera toxin subunit B. (B–C) Densitometric quantification of E-cadherin (B) and γ-catenin (C) distribution in the fractions. The signals were normalized to fraction number 12. Data points represent the mean ± SD of three independent experiments. Two-way ANOVA with Bonferroni’s multiple comparison test. *, p<0.05; **, p<0.01; ***, p<0.001.