Fig. 1A–C.

Effect of α-TOS on tumor cell viability in vitro. Lymphocytes, dendritic cells or 3LL cells were plated in 6-well tissue culture plates. A The cells were treated in triplicate with 10, 20 or 40 μg/ml α-TOS or 0.01% ethanol. After 24 h, nonadherent and adherent cells were collected and evaluated for cell viability using AO and PI. B To evaluate cell yield, 3LL cells were treated with 10, 20, 40 or 80 μg/ml α-TOS, sodium succinate (NaS), or 0.01% ethanol (E). At each time point (4 h, 8 h, 24 h), nonadherent and adherent cells were collected and cell yield determined. No significant effect was observed at the 4-h or 8-h time point (data not shown). The results (A, B) depict the mean ± SEM of three independent experiments. C For the apoptosis assay, cells were treated with either 40 μg/ml α-TOS or NaS. At each time point, nonadherent and adherent cells were collected and stained using annexin V and PI. Numbers represent the percentages of cells with phosphatidyl serine externalization (lower right quadrant) and cells with loss of membrane integrity (upper right quadrant), respectively