Oxaprozin-Induced Apoptosis on CD40 Ligand-Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways
Figure 3
Oxaprozin-induced apoptosis involves the inhibition of CD40L-induced Akt and phosphorylation. Monocytes were incubated for 48 hours with control medium or 200 ng/mL CD40L plus 1 g/mL CD40L enhancer in the presence or absence of 50 M LY294002 (LY), 50 M PD98059 (PD), 1 M SB203580 (SB), or 20 M SN-50. (a) Assessment of apoptosis on cytopreps stained with acridine orange. Data are expressed as percentages of apoptotic cells on total number of cells counted (mean SEM, n 3, apoptosis in absence versus in presence of CD40L: **P .01; apoptosis in presence of CD40L versus CD40L plus PD: ##P .01; apoptosis in presence of CD40L versus CD40L plus LY or SB or SN-50:#P .05). (b) Assessment of apoptosis using Annexin V and Propidium Iodide (PI) binding assay. Data are expressed as percentages of positive cells on total number of cells counted (mean SEM, n 2). (c) Monocytes were prencubated in the presence or absence of 100 M oxaprozin for 1 hour and then stimulated with control medium or 200 ng/mL CD40L plus 1 g/mL CD40L enhancer for 15 minutes. Then, cells were lysed, and western blot analysis was performed. Results represent one of three experiments that yielded similar results.
