A: MOLT-14 cells were treated with PBS or APP in cRPMI for the time indicated in the figure. Cell lysates were prepared and subjected to Western immunoblot for phospho-ERK1/2 or total ERK1/2. Data are representative of three independent experiments. B: Cells were treated with APP, APP and 20μM U0126, or mock and harvested at 15, 30, 60, and 240 min. Cells were then lysed and analyzed for phospho-ERK1/2 or ERK1/2 via western blot. C: MOLT-14 cells were treated with PBS or APP in cRPMI for two hours, followed by treatment with U0126 for two hours, and then with actinomycin D for the time indicated in the figure. RNA was extracted from each sample and subjected to quantitative RT-PCR for 18S, GM-CSF, and IL-8. Data represent mean of triplicate values +/− SD and are presented as the normalized amount of each transcript to 18S. The percentage of each transcript that remained following treatment was calculated after subtracting background (PBS t=90m). Data are representative of three independent experiments.