Induction of apoptosis in C4-2B cells by MK591 treatment.

<p>C4-2B cells (~3 x 10⁵ per plate) were plated in 60 mm diameter plates and allowed to grow for 48 hours. Then the old medium was replaced with 2 ml fresh RPMI medium and the cells were treated either with MK591 or ibuprofen for 24 hours at 37^°C. Control cells were treated with vehicle only (0.2% DMSO). In (a), at the end of incubation period, intact cells were treated with FITC-labeled Annexin-V and propidium iodide (PI) in binding buffer and observed under microscope. Photographs were taken with a Nikon digital camera attached to a Leica fluorescence microscope at x200 and processed on a Dell computer using Q-Capture Pro7 software. In (b and c), at the end of incubation period, cells were lysed, and cleavage of PARP (c-PARP) and caspase-3 were detected by Western blot. In (d), degradation of nuclear DNA was detected by Cell Death Detection ELISA. Results are shown as mean values of each data point ± standard error (**p < 0.005, n = 4).