Fractionation of an Extract of Pluchea odorata Separates a Property Indicative for the Induction of Cell Plasticity from One That Inhibits a Neoplastic Phenotype
Figure 1
Antiproliferative effect of (a) dichloromethane crude extract, (b) fraction F1.1 in HL60 cells; 1 × 105 cells/mL were seeded into 24-well plate, incubated with 5, 15, and 40 μg/mL extract or fraction F1.1, and the percentage of proliferation was calculated relative to solvent treated control within a 24 h period, (c) Induction of apoptosis by F1.1: cells were grown as described and incubated with 15 and 40 μg/mL of each fraction for 72 h. Then, cells were double stained with Hoechst 33258 and propidium iodide and examined under the microscope with UV light connected to a DAPI filter. Nuclei with morphological changes which indicated cell death were counted, and the percentages of dead cells were calculated. Experiments were performed in triplicate. Asterisks indicate significance compared to untreated control (), and error bars indicate ±SD. (d) 1 × 106 cells/mL were incubated with 40 μg/mL F1.1 and harvested after 0.5, 2, 4, 8, and 24 h of treatment, lysed, and total protein applied to SDS-PAGE. Western blot analysis was conducted with the indicated antibodies. Equal sample loading was confirmed by Ponceau S staining and β-actin analysis.
