Figure 1.
Effect of EPSAH on the cell proliferation in human HeLa cells.
HeLa cells were seeded in 96-well culture plates. After incubation for 24, 48 or 72 h, they were subjected to MTT assay. Results are expressed as percent cell proliferation relative to the proliferation of control. Data represents as mean ± S.D. from three independent experiments. *P<0.01 vs control.
Figure 2.
Morphological changes in HeLa cells treated with different concentrations of EPSAH (50–800 µg/ml) for 48 h and compared with untreated cells.
The cells were examined under the light microscope after Wright-Giemsa stain. Nuclear fragmentation, chromatin condensation and/or chromatin margination was observed in EPSAH-treated HeLa cells. The most representative fields are shown.
Figure 3.
Identification of apoptotic cells by AO/EB staining.
HeLa cells were treated with different concentrations of EPSAH (50–800 µg/ml) for 48 h and compared with untreated cells. The most representative fields are shown.
Figure 4.
Agarose gel etectrophoresis of DNA from HeLa cells treated for 48 hours with different concentrations of EPSAH.
Figure 5.
Effect of EPSAH on the level of the cleaved caspase-3 in HeLa cells.
The level of the cleaved caspase-3 were assayed by Western blotting in the cells treated with EPSAH at the indicated doses for 48 h.
Figure 6.
EPSAH-induced mitochondrial transmembrane potential in HeLa cells measured by Rh123 staining and analyzed by flow cytometry.
Figure 7.
EPSAH induced Bcl-2, Bax, P53, survivin, Grp78 and CHOP protein expression levels in HeLa cells.
Cells were treated with EPSAH at the concentrations of 100 and 400 µg/ml for 48 h. The treated and untreated cells were lysed and cell extract subjected to western blots with the antibodies as described in the Methods section. Immunoblot images are the representative of 3 identical experiments. The GAPDH blot is a loading control. (A) Bax/Bcl-2 proteins, (B) P53 and survivin proteins (C) Grp78/CHOP proteins.
Figure 8.
A potential mechanism of EPSAH-induced cell apoptosis in HeLa cells.
(↑) and (↓) arrows indicate increase and decrease.