10.1371/journal.pone.0107002.g003 Justin L. Eddy Justin L. Eddy Lindsay M. Gielda Lindsay M. Gielda Adam J. Caulfield Adam J. Caulfield Stephanie M. Rangel Stephanie M. Rangel Wyndham W. Lathem Wyndham W. Lathem OMVs contain outer membrane-associated virulence factors. Public Library of Science 2014 protease Pla Yersinia pestis membrane vesicles plasminogen activation fimbrial antigen Braun lipoprotein Membrane proteins Plague Pathogen Yersinia pestis Virulence factors Pla substrates Fas ligand F 1 membrane stress disease plague pestis OMVs infection OMV release adhesin Ail Outer Membrane Vesicles Cell growth Extracellular matrix 100 nm 2014-09-08 03:16:10 Figure https://plos.figshare.com/articles/figure/_OMVs_contain_outer_membrane_associated_virulence_factors_/1165090 <p>(A) Whole cell lysates (WCL) or density centrifugation gradient fractions from OMVs isolated from <i>Y. pestis</i> were separated by SDS-PAGE and gels were silver stained. (♦) denotes an enriched band and (•) denotes reduced bands. (B) WCL or gradient fractions (4–6) from OMVs isolated from <i>Y. pestis</i> and <i>Y. pestis</i> Δ<i>pla</i> were examined for the presence of the virulence factors Pla, Ail, and Caf1 by immunoblot. Immunoblots for RpoA and Hfq, two cytoplasmic proteins, are shown to demonstrate the absence of contaminating proteins from the OMV preparation. (C) OMVs were immuno-gold labeled with an anti-Caf1 antibody conjugated to gold beads and examined by TEM. Black arrows indicate representative gold particles. Bar represents 50 nm. (D) Subcellular distribution of proteins present in <i>Y. pestis</i> OMVs as a percentage of total proteins identified by mass spectrometry listed in (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107002#pone.0107002.s003" target="_blank">Table S3</a>).</p>