Supporting information for Giannakakou et al. (2002) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.132275599.
Fig. 9.
(A) Effect of PTX and adriamycin (ADR) combination on cell survival. A549 cells were plated in 96-well plates in triplicate and treated with a fixed concentration of PTX at 3 nM, or 6 nM for 24 hr followed by 400 ng/ml of ADR for another 4 days. Cells treated with PTX or ADR alone are included as controls. Cell survival was determined with the sulforhodamine-B assay, and the OD564 was measured in a microplate reader. The bar graph represents percent survival as determined by the OD value of the treated cells divided by the OD value of untreated cells. The mean OD values were determined from three independent experiments. Error bars represent standard error of the mean. (B) Effect of PTX and ADR combination on caspase-3 activation. A549 cells were plated in 96-well plates in sextuplet and treated with 3 nM or 6 nM PTX overnight followed by 400 ng/ml of ADR for an additional 6 hr. Untreated cells or cells treated with ADR or PTX alone are included as controls. After drug treatment, cells were tested for caspase-3 activity by the HTS Caspase-3 Activity Fluorometric Assay (Oncogene Science), according to the manufacturer’s instructions. The values of the y axis (random fluorescent units) represent the mean of two independent experiments where cells were plated in sextuplet each time (mean of 12 values). Error bars represent SEM. Control is arbitrarily assigned the value of 10, and the data presented are normalized to the value of the control.