Fig. 7.
Adriamycin-induced apoptosis in wild type cells and MPGES-1 knockdown clones. (A) Apoptosis assay. Cells (7 × 103) were seeded on 96-well plates and allowed to attach for 24 h. New medium containing 300 nM adriamycin was added, and after 24 h cells were lysed, and cytokeratin 18 fragments were analyzed by ELISA. For each condition, two incubations were analyzed (n = 2), and results are given ± SD. (B) Western blot analysis of caspase-3 fragment in DU145 cells—WT and MPGES-1 knockdown clone A—after treatment with adriamycin (300 nM) for 24 h. Cells were lysed with M-PER buffer, and 50-μg aliquots of total protein were applied to the SDS/PAGE gel. β-Actin analysis shows equal loading. After electroblotting, the membrane was incubated with an antibody for caspase-3 cleaved at Asp-175.