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. 2013 Jun 14;8(6):e66244. doi: 10.1371/journal.pone.0066244

Figure 4. SEG co-localizes with MHC-II at the cell membrane and in vesicles.

Figure 4

DCs were cultured in poly-L-lysine treated slides, pulsed 1 h at 37°C with SEG-FITC, washed four times and cultured for different periods (0–240 min). Cells were then fixed with paraformaldehyde and permeabilized with saponin for immunolabeling with a phycoerythrin-Cy5-labeled monoclonal antibody against mouse I-A and I-E MHC-II. A control of non-pulsed cells was performed in parallel. Confocal images were captured using a PlanApo 60× Oil AN1.40 lens. After 1 h pulse, SEG showed a dispersed pattern, with some clusters, near the cell membrane. Strong staining of MHC-II was observed at a similar location. At 60 min, SEG co-localized with MHC-II in vesicles. One hundred and eighty min later, clusters of MHC-II and SEG were observed, in addition to a scattered pattern for both SEG and MHC-II close to the cell membrane. Figure shows a representative experiment of 5.