Expression of LYVE-1 on LECs.

<p>SVEC4-10, a SV40-immortalized endothelial cell line derived from mouse axillary lymph nodes, is a LEC-like cell line. (<b>A</b>)The expression of LYVE-1 on SVEC4-10 was assessed by immunofluorescence staining, confocal microscopic analysis and western bloting. Cells were grown on cover slips and fixed with 4% paraformaldehyde for 10 min. SVEC4-10 cells were stained with PBS (a) or anti-LYVE-1 mAb (b), followed by Alexa Fluxo 488-conjugated goat anti-rat IgG and visualized with a confocal microscope. COS-7^{LYVE−1 (+)} (d) and COS-7^pERFP−N1 (c) were included as the positive control and negative control, respectively. The white arrows indicate the plasma membrane localization of LYVE-1. Magnification was ×600. (<b>B</b>) In addition, the cell lysates of SVEC4-10, COS-7^{LYVE−1 (+)} and COS-7^pERFP−N1 were subjected to 7.5% SDS-PAGE followed by Western blot analysis using antibody to LYVE-1. The closed and opened arrows indicate the locations of the monomer and dimer of LYVE-1, respectively. The arrowhead indicates the location of a proteolytic product of LYVE-1. The slight difference of the location of LYVE-1 between COS-7^{LYVE−1 (+)} and SVEC4-10 cells may due to cell type-specific glycosylation <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092857#pone.0092857-Nightingale1" target="_blank">[49]</a>. Total GAPDH was used for normalization. Data were representative of three independent experiments.