Figure 3

A. Nuclear Morphology of Jurkat cells treated with AMD5 at 10 μM for either 24 hrs or 48 hrs. Jurkat cells were stained with cell-permeable Hoechst 33342 dye to observe apoptotic nuclear morphology. Apoptotic nuclei are brightly stained and condensed when compared to healthy nuclei. The control was treated with DMSO solvent. Magnification: 200×. B. Annexin-V Binding Assay: Jurkat cells were treated with 10 μM AMD5 for 24 hrs. Jurkat cells were incubated with Annexin-V 488 Alexa-Fluor conjugate to observe phosphatidyl-serine flipping from the inner to the outer leaf of the plasma membrane. This is a characteristic event of apoptosis that is made visible with use of a fluorescent microscope; cells undergoing apoptosis will have more Annexin-V substrate bound, and thus appear brighter than healthy cells. Magnification: 400×