mtMPs were detected in plasma from FPI mice and contained surface-exposed CL. Transmission electron microscope images. (A) Intact (left) and membrane-disrupted (middle, arrow) mitochondria and mitochondria-embedded BDMP (right; asterisk, mitochondria) detected in plasma of FPI mice. (B) Mitochondria (asterisk), membrane microparticles (arrowhead), and (C) mitochondria-embedded membrane microparticles (*) from purified BDMPs. (D) A section of an uninjured mouse brain shows a dense perinuclear distribution of mitochondria (left, arrowhead: nuclear membrane). A locally enlarged image further shows mitochondria (asterisk) and endoplasmic reticulum (ER) and membrane-bound/free polyribosomes (arrow). (E) Total mtMPs and (F) mtDNA detected in plasma samples from sham and FPI mice. (G) Annexin V bound to more than 80% of mtMPs purified by TOM22 antibody (left), and the binding was abolished by EDTA (right). (H) Anti-CL antibody purified from FPI mice, but not a control IgG, bound purified mtMPs. (I, top) CL and PS captured to the nitrocellulose membrane were incubated with human annexin V (PBS and annexin V as controls). Annexin V bound to the phospholipids was detected by a polyclonal annexin V antibody. (Bottom) Allophycocyanin-conjugated annexin V bound to CL and PS coated on microbeads detected by flow cytometry (n = 3-6/group; paired Student t test).