Figure 1.
Chemical structure of dammarane skeleton of G. pentaphyllum.
Figure 2.
Effects of Gyp on cell viability.
SW-480 were seeded in 96-well plates and treated with 0, 70, 100 and 130 µg/ml of Gyp for 24 and 48 h. Cell viability was measured by MTT assay. Each value is expressed as a mean ± S.D. of at least three independent determinations. One-way ANOVA was used for comparisons of multiple group means followed by Dunnett’s t-test. **P<0.01 versus the control. (error bars = S.D., n = 3).
Figure 3.
Effects of Gyp on the cell membrane integrity of SW-480 cells.
Cells were treated with Gyp for 6, 12, 24 and 48(vertical axis) vs. PI fluorescence (horizontal axis).
Figure 4.
Effects of Gyp on mitochondrial membrane potential (Δψm) of SW-480 cells.
Cells were treated with Gyp for 4, 8 h and labeled with rhodamine 123 and analyzed by flow cytometry. Histograms show number of cell channel (vertical axis) vs. rhodamine 123 fluorescence (horizontal axis).
Figure 5.
Effects of Gyp on DNA fragmentation of SW-480 cells.
Cells were treated with Gyp for 6, 12, 24 and 48(vertical axis) vs. PI fluorescence (horizontal axis).
Figure 6.
Gyp effects on the cell morphology and nucleus in SW-480 cells.
Cells were treated with Gyp at the indicated concentrations for 6, 24, 48“Materials and methods”.
Figure 7.
Apoptosis-inducing effect of Gyp on SW-480 cells.
Dot plots of Annexin V and 7-AAD uptake after indicated concentrations in SW-480 cells. Cells were analyzed at 12, 24 h post treatment.
Figure 8.
Effects of Gyp on the migration of SW-480 cells in vitro.
Cells in 24-well plates were wounded by scratching with a pipette tip and the cells were incubated with Gyp for 24 hours. The cells were photographed under phase-contrast microscopy (×200 magnification).
Figure 9.
Changes in the actin cytoskeleton of SW-480 cells after Gyp.
Cells were stained with phalloidin (green) for F-actin. Scale bars, 10 µm.
Figure 10.
Scanning electron microscopic images of SW-480 cells at 24 h after Gyp.
Magnification of the images was 1500 and 5000×, respectively. Scale bars: 30, 10 µm.
Figure 11.
Gyp-induced production of reactive oxygen species (ROS) in SW-480 cells.
Cells were treated with Gyp for 4, 8–DA and the fluorescence intensity of the oxidized product DCF in individual cells was detected by flow cytometry. The percentage of fluorescent cells in each group was shown.
Figure 12.
Effect of ROS scavenger NAC (N-acetylcysteine) on the effect of Gyp on SW-480 cells.
(A) Cell viability, (B) intracellular ROS generation, (C) nuclear morphological changes, (D) cell migration, (E) Δψm loss were specially analyzed as described in “Materials and methods”.