Fig. 4

Subcellular localization of Lip40. a SDS-PAGE analysis of A. pleuropneumoniae subcellular fractions. b Western blotting analysis. Subcellular fractions were separated with 12 % SDS-PAGE, blotted onto nitrocellulose membranes, and incubated with rabbit anti-rLip40 polyclonal antibody. Color was developed with a DAB peroxidase substrate (Tiangen). M, prestained protein ladder; lane 1, cytoplasmic proteins (CP); lane 2, periplasmic proteins (PP); lane 3, cytoplasmic membrane proteins (CM); lane 4, outer-membrane proteins (OM); lane 5, extracellular proteins (EP). The nitrocellulose membranes displayed a band with a molecular weight of ~30 kDa in the outer-membrane fraction, indicating that Lip40 is located on the outer membrane. No signal was detected in the other subcellular fractions