Figure 2

Anti-proliferative property of AB215 on ERα ⁺ breast cancer cells. A, B) ERα⁺: MCF7, C, D) T47D and E, F) ERα⁻: SK-BR-3, cells were grown in phenol red free RPMI1640 supplemented with 2% heat inactivated charcoal-stripped FBS treated with or without 10nM E2 along with 500 ng/ml BMP2 or AB215 in quintuplicate. Cell proliferation was analyzed by MTT assay (Abs590-700 nm) on 0, 1, 3 and 5 days after the treatment (n = 3). E2 and AB215 did not affect the proliferation of ERα⁻ cells significantly. The results are presented as means ± SD and their significance has been analyzed by one-way ANOVA. G) MTT assay of MCF7 and T47D cells at increasing concentration of AB215 in the presence of 10nM E2. Cells were plated and treated as explained in Figure 2a-f. Cells were analyzed 4 days after the treatment. Data are shown in means + SD. H) Western blot analysis of E2 induced phosphorylation of ERK1/2. Cells were plated in phenol red free RPMI1640 supplemented with 5% heat inactivated charcoal-stripped FBS treated with or without 10nM E2 along with 500 ng/ml BMP2 or AB215. Cells were harvested and lysed after 48 hours of exposure for western blot analysis. (* = P ≤ 0.05, ** = P ≤ 0.01, *** = P ≤ 0.001).