Phyllanthus urinaria Induces Apoptosis in Human Osteosarcoma 143B Cells via Activation of Fas/FasL- and Mitochondria-Mediated Pathways
Figure 2
Apoptosis of 143B and 143Bρ 0 cells of P. urinaria detected by flow cytometry with annexin V and propidium iodide (PI) and TUNEL staining. 143B and 143Bρ 0 cells treated with P. urinaria for 24 h (a) and 48 h (b) are shown with representative dot plots from FITC-conjugated annexin V and PI staining. Early apoptosis was recognized in the presence of staining for annexin V and absence of staining for PI. Late apoptosis was recognized in the presence of staining for both annexin V and PI. The percentage of apoptosis from flow cytometric analysis for 24 h (c) and 48 h (d) was assessed. Values are mean ± SD of three independent experiments in duplicate. Asterisks represent statistical significance from the vehicle control (*, **). A pound sign represents statistical significance between 143B and 143 Bρ 0 cells (#). Immunofluorescence showing apoptotic 143B and 143Bρ 0 cells marked by TUNEL assay in the absence and presence of 2 mg/mL P. urinaria treatment for 24 h (e) and 48 h (f) are indicated. DAPI staining was used to detect all nuclei and visualized under a fluorescence microscope (1000x). DNA damage of 143B cells in comparison with 143 Bρ 0 cells was markedly increased with the treatment of P. urinaria (#).
