Figure 6 | Scientific Reports

Figure 6

From: Microvesicles released from pneumolysin-stimulated lung epithelial cells carry mitochondrial cargo and suppress neutrophil oxidative burst

Figure 6

Pneumolysin (PLY) triggers the release of microvesicles (MVs) carrying mitochondrial cargo. (A) MitoTracker Deep Red and Green staining of A549 cells after treatment with PLY (100 ng/ml, 4 h) or PBS (Ctr). Gated cells stain less brightly with Mitotracker Deep Red after PLY indicative of loss of membrane potential. (B,C) MitoTracker Deep Red-labeled A549 were challenged with PLY (100 ng/ml, 4 h) or PBS. MVs were analyzed by FACS (B) or after staining with (C) CFSE or annexin V-PE. (D–F) A549 were challenged with (D) PLY (100 ng/ml, 4 h), (E) Spn WT vs Δply (MOI 50, 4 h), or (F) pre-treated with MitoTempo (50 μM) or DMSO prior to PLY (100 ng/ml, 4 h). MVs were isolated from the conditioned media and analyzed by immunoblotting. Depicted are representative blots of Tom20 (mitochondrial protein) and pan-cytokeratin (cropped images). Experiments were performed 3 or more independent times.

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