Figure 4

FW-04-806 inhibits growth, induces cell cycle arrest, induces apoptosis, and downregulates the expression of anti-apoptotic proteins. (A) SKBR3 and MCF-7 cells were grown for 48 h in the absence or presence of increasing concentrations of FW-04-806. Cell growth inhibition was measured with MTS and was expressed as a percentage of vehicle-treated control; results are presented as means ± SD of three independent experiments. *p < 0.05: significant difference from control by analysis of variance; **p < 0.01: very significant difference from control by analysis of variance. (B) SKBR3 and MCF-7 cells were treated with increasing doses of FW-04-806 or vehicle DMSO for 24 h. The cells were then fixed with 70% ethanol at −20°C overnight, incubated with RNase A at 37°C for 30 min, stained with propidium iodide for 10 min, and analyzed with flow cytometry. (C) SKBR3 and MCF-7 cells were treated with FW-04-806 for 24 h, and apoptotic cell death was detected by staining cells with an Annexin-V: FITC Apoptosis Detection Kit for analysis with flow cytometry. (D) SKBR3 and MCF-7 cells were treated with increasing doses of FW-04-806 for 24 h, and the apoptosis signal proteins were detected with western blot analysis using an Apoptosis Antibody Sampler Kit; β-actin was used as a loading control.