Fig. 5

Lunasin increases E-cadherin expression in mouse HC11 mammary epithelial cells and does not affect mammosphere formation of MCF-7 cells. Non-nuclear (a) and nuclear (b) extracts were isolated from HC11 cells treated with vehicle or 1 μM Lunasin (LUN) for 6 h and analyzed for E-cadherin and β-catenin proteins by western blots. Loading controls used were α-tubulin (a) and Lamin B1 (b). Representative western blots are shown, with each lane representing samples from an independent experiment. Densitometric scans are normalized to α-tubulin or Lamin B1 (mean ± SEM). *Significant difference at P < 0.05, using Student’s t test. c Transcript levels for C-MYC in HC11 cells treated with vehicle or LUN (1 μM) with and without added recombinant Wnt1 (10 ng/ml) were quantified by qRT-PCR, Values were normalized to those of corresponding 18S and renormalized to vehicle-treated group. Means (±SEM) that were expressed as fold-change are from three independent experiments, with each experiment performed in triplicate. Different superscripts indicate significant differences (P < 0.05, one-way ANOVA). d MCF-7 cells were treated with vehicle, 2 μM LUN or 2 μM LUN + 40 nM genistein (LUN + GEN) in low-attachment plates. Primary (P1) mammospheres were collected at day 5 and passaged for secondary (P2) mammospheres without additional treatments. Mammosphere formation at P2 was evaluated at day 7 post-plating. Mean MFUs (±SEM), relative to vehicle-only-treated cells, are from P2 mammospheres of two independent experiments, with each experiment performed in quadruplicate. Means with different superscripts are significantly different (P < 0.05, one-way ANOVA)