Staphylococcus aureus Panton-Valentine leukocidin directly targets mitochondria and induces Bax-independent apoptosis of human neutrophils
J. Clin. Invest. Anne-Laure Genestier, et al. 115:3117 doi:10.1172/JCI22684 [Go to this article.]

Figure 6
rPVL is localized in mitochondria during rPVL-induced PMN apoptosis. (A) PMNs were treated for 5 minutes with or without 5 nM biotinylated rPVL and were then stained with streptavidin-FITC before (without permeabilization) or after (permeabilization) treatment with Cytofix/Cytoperm. (B) PMNs were incubated with medium or with 5 nM PVL and, at the indicated times, cytosolic and mitochondrial protein extracts were prepared and proteins were separated on 12% SDS-PAGE followed by Western blotting with anti-MnSOD, anti–β-actin, anti–LukS-PV, or anti-Bax Ab. Anti-MnSOD Ab served as a marker of mitochondrial contamination of the cytosolic fractions (Cyto), while anti–β-actin Ab served as a marker of cytosolic contamination of mitochondrial fractions (Mito). Data are from 1 of 2 independent experiments that gave similar results. (C) PMNs were treated for 5 minutes with or without 5 nM biotinylated rPVL, prepared for TEM, and stained with gold-conjugated streptavidin. Arrows indicate gold-positive cells. Scale bars: 500 nm.