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. 2015 Feb 26;19(5):1133–1150. doi: 10.1111/jcmm.12518

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© 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Transmission electron microscopy (TEM) and confocal immunofluorescence imaging of mitochondrial remodelling in BMSCs challenged with Staphylococcus epidermidis. TEM micrographs and projections of Tom 20 obtained from control BMSC (A) and BMSCs fixed at 5 hrs (B) and 24 hrs (C) following the bacterial challenge. The images shown in the figure are representative ones of analyses of three specimens obtained from three experiments. (A1, B1, B3 and C1) Low-magnification TEM micrographs of BMSCs. (A2) High-magnification TEM micrograph of normal mitochondrion. (B1) Aberrant mitochondria and formation of mitophagosome are indicated with white arrows. (B2) High-magnification TEM micrograph of a swollen (aberrant) mitochondrion with lysis of cristae and transfer of an aberrant mitochondrion to autophagosome (indicated with white arrows). (B3) Co-localization of xenophagy of S. epidermidis (S.e.) and mitophagy events in BMSC. Aberrant mitochondria and formation of mitophagosome are indicated with white arrows, S. epidermidis in an autolysosome is shown with a white arrowhead. (B4) (ROI indicated in B1). Image of aberrant mitochondria in autolysosome. (B5 and B6) Autophagy of aberrant mitochondria (mitophagosome formation). (B7) Remodelling of aberrant mitochondria as indicated with black arrows. (C1 and C2) TEM images of mitochondrial alterations observed 24 hrs after bacterial challenge. Formation of fragmented aberrant mitochondria (white arrows) and elongated mitochondrial tubules (black arrows) is shown in C1. Fusions of aberrant mitochondria with autophagosomes are indicated in C2 with white arrows. (A3, B8 and C3) Confocal immunofluorescence images of mitochondrial remodelling in BMSCs. The mitochondrial networks were visualized using projections of Tom20 (red channel), a mitochondrial marker. (A3) Control BMSCs. The mitochondrial network is presented by a combination of elongated and round-shape projections. (B8) Mitochondrial alterations observed 5 hrs after bacterial challenge. The mitochondrial network is comprised of numerous round-shape projections. (C3) Mitochondrial alterations observed 24 hrs after bacterial challenge. The mitochondrial network appeared in a form of reticular long-length structures formed because of mitochondrial fusion. TEM conditions: Same as indicated in Figure2. The confocal images were taken with pinhole setup to obtain 0.5 μm Z-sections.