SDS-PAGE, immunoblot analyses, Nanoparticle tracking analysis and electron microscopic analyses of OMV samples from the wild type strain C6706 and the prtV mutant.

(A) SDS-PAGE and Coomassie blue staining of OMVs samples from V. cholerae wild type C6706 (lane 1) and its derivative prtV mutant (lane 2). (B) Immunoblot analysis of PrtV protein in the OMV samples from the wild type strain C6706 (upper panel, lane 1) and the prtV mutant (upper panel, lane 2) using anti-PrtV polyclonal antiserum. Immunoblot analysis of OMV samples using anti-OmpU antiserum as a OMV marker (middle panel) and anti-Crp polyclonal antiserum as a cytoplasmic protein marker (lower panel). (C) Nanoparticle tracking analysis measurement of OMVs isolated from the wild type V. cholerae strain C6706 showing the sizes and total concentration of OMVs. (D) Nanoparticle tracking analysis measurement of OMVs isolated from the ΔprtV mutant showing the sizes and total concentration of OMVs. (E) Electron microscopy of OMVs from the wild type V. cholerae strain C6706 (a) and the prtV mutant (b). Immunogold labeling of OMVs from V. cholerae wild type strain C6706 (c) and the prtV mutant (d). White arrow points to gold particles associated with OMVs. Bars; 150 nm.