FIG. 6.

p38-MAPK activation levels in CCFSSTG1 and U87MG cells treated with biochanin A (BCA) and other phytoestrogens/steroids in a prevention regimen. Representative Western blots of phospho-p38-MAPK (Thr180/Tyr182) and total p38-MAPK levels in CCFSTTG1 (A) and U87MG (B) cells treated with 0, 10, 20, 40, 60, 80, and 100 μM BCA or genistein (GEN), or 0, 10, 20, 40, 60, 80, and 100 nM dexamethasone (DEX) or β-estradiol (E2) for 24 h, followed by treatment with ibuprofen (2 mM) for 24 h followed by treatment with ibuprofen (2 mM) + 0, 10, 20, 40, 60, 80, and 100 μM BCA, genistein or 0, 10, 20, 40, 60, 80, and 100 nM dexamethasone (DEX) or β-estradiol for an additional 24 h. Cells were lysed after treatment completion and equal amounts of protein (40 μg) were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblots were stripped and reprobed for total p38-MAPK levels. Quantification of the mean of phospho-p38-MAPK/p38-MAPK ratio levels from two independent experiments, respectively, is shown on the right. Values are normalized to levels in ibuprofen-treated cells only.