Subcellular redistribution of Bax protein in PMNs.

PMNs were cultured for 6 hours without additions (No stimulus), with 20 ng/mL TNF-α, or with a combination of 20 ng/mL TNF-α and 150 μM zVAD-fmk. Then the cells were stained with MitoTracker GreenFM, fixed, permeabilized, stained with polyclonal Abs specific for Bax, and analyzed with CLSM. (Because of the fixation and permeabilization procedures, the mitochondrial staining [green] showed a more diffuse cytoplasmic pattern than the tubular structures shown in Figure 4, left panels). Each image represents the following proportion of the total cell population (mean ± SEM): 77.7% ± 5.4% in No stimulus; 63.3% ± 8.8% in TNF-α; 88.3% ± 5.7% in TNF-α/zVAD-fmk. Bar is 5 μm. This figure is representative of at least 4 independent experiments.