Figure 1. . Quercetin and 2-methoxyestradiol inhibit cell proliferation in a time-dependent manner with simultaneous synergistic inhibition of TGF-β-induced migration and colony-forming ability of PC-3 cells.

(A) Effect of drug treatment on PC-3 cell proliferation was analyzed by MTT assay, normalized with the control group and graph was plotted with OD at 540 nm on Y-axis and time on X-axis. Data represented as mean ± SEM from three independent experiments. The statistical analysis was performed using ANOVA with Tukey’s post hoc multicomparison test (*p < 0.05; **p < 0.01; ***p < 0.001). (B) Cell migration in scratch assay after treatment with all the three groups of drugs. Pictures were taken using an inverted microscope in various time points in triplicates. Data represented as mean ± SEM from three independent experiments. (C) Histogram plots with the percentage migration in the respective treatment group are shown as mean ± SEM from three independent experiments. The statistical analysis was performed using ANOVA with Tukey’s post hoc multicomparison test (*p < 0.05; **p < 0.01; ***p < 0.001). (D) Soft agar assay was performed to determine the colony-forming ability of PC-3 cells. PC-3 cells were harvested after 24 h of treatment and suspended in serum-free medium and plated in soft agar dishes prepared using 1% base agar and 0.7% top agar layers. Pictures were taken every 24 h using an inverted microscope to observe the transformation of cells.

2-ME: 2-methoxyestradiol; EMT: Epithelial–mesenchymal transition; OD: Optical density; Qu: Quercetin.