Table 2.
Apoptosis detection by annexin V binding/PI uptake assay in SiHa parental and SiHa CDV
| |
SiHa
parental
|
SiHa
CDV
|
||||||
|---|---|---|---|---|---|---|---|---|
| Untreated | CDV 158.7 μM | CDV 63.5 μM | PMEG 6.5 μM | Untreated | CDV 158.7 μM | CDV 63.5 μM | PMEG 6.5 μM | |
| Early apoptotic |
1.2 ± 0.9 |
22.1 ± 0.6 |
8.4 ± 1.6 |
64.2 ± 1.2 |
4.5 ± 2.0 |
2.3 ± 0.1 |
2.4 ± 0.2 |
30.6 ± 4.3 |
| Late apoptotic |
0.6 ± 0.2 |
5.0 ± 5.6 |
2.7 ± 2.5 |
6.0 ± 4.2 |
0.9 ± 0.4 |
0.6 ± 0.1 |
0.6 ± 0.1 |
3.9 ± 0.7 |
| Viable |
97.2 ± 2.2 |
72.2 ± 6.6 |
88.0 ± 3.9 |
29.4 ± 3.0 |
94.5 ± 2.2 |
96.3 ± 0.5 |
96.5 ± 0.3 |
64.9 ± 1.2 |
| Necrotic | 0.4 ± 0.3 | 0.7 ± 0.3 | 0.9 ± 0.2 | 0.4 ± 0.1 | 0.2 ± 0.1 | 0.8 ± 0.6 | 0.5 ± 0.2 | 0.7 ± 0.3 |
Cells were seeded in 6-well microtiter plates at a density of 5 × 104 cells/well and allowed to proliferate for 24 h. Culture medium was removed and medium containing the compounds was added. At 7 days post-treatment, cells were harvested with EDTA/PBS (760 mg/L). After rinsing the cells with PBS, the cell pellets were simultaneously stained with annexin-FITC and PI using the annexin-V-FITC staining kit (BD Biosciences, San Jose CA) and analyzed by flow cytometry. Dual fluorescence dot plots were obtained following combined annexin V-FITC and PI staining. Data represent the percentages in each quadrant of the dot plots and are the mean ± SD of at least two stainings. Viable (annexin V-/PI-) early apoptotic (annexin V+/PI-), necrotic (annexin V-/PI+), and late apoptotic (annexin V+/PI+) cells.