Skip to main content
. 2006 Jul;80(13):6305–6317. doi: 10.1128/JVI.02545-05

FIG. 7.

FIG. 7.

Analysis of ZEBOV- and SEBOV-GP glycosylation. (A) lVLPs were produced by transient transfection of 293T cells in the presence or absence of DMJ, normalized for comparable Gag contents, digested with endo H and PNGase F for the indicated time periods, and analyzed by Western blotting. (B) lVLPs bearing the indicated GPs were concentrated by centrifugation through a sucrose cushion and analyzed for GP and Gag incorporation by Western blotting with antisera raised against ZEBOV-GP and Gag, respectively. lVLPs were obtained from cells expressing the following proteins: lane 1, Gag; lane 2, Gag and SEBOV-GP; lane 3, Gag and ZEBOV-GP; lane 4, Gag and S33Z variant; lane 5, Gag and Z33S variant. (C) lVLPs were analyzed for reactivity with lectins with known specificities. lVLPs were derived from cells expressing the following proteins: lane 1, Gag; lane 2, Gag and SEBOV-GP; lane 3, Gag and ZEBOV-GP; lane 4, Gag and S33Z variant; lane 5, Gag and Z33S variant. (D) Supernatants from pcDNA3 (lanes 1)-, SEBOV-GP (lanes 2)-, and Z33S (lanes 3)-transfected cells were concentrated using Centricon Plus columns and analyzed as described for panel C. Panels C and D show the results of representative experiments, and similar results were obtained in two independent experiments. cushion and analyzed for GP and Gag incorporation by Western blotting with antisera raised against ZEBOV-GP and Gag, respectively. lVLPs were obtained from cells expressing the following proteins: lane 1, Gag; lane 2, Gag and SEBOV-GP; lane 3, Gag and ZEBOV-GP; lane 4, Gag and S33Z variant; lane 5, Gag and Z33S variant. (C) lVLPs were analyzed for reactivity with lectins with known specificities. lVLPs were derived from cells expressing the following proteins: lane 1, Gag; lane 2, Gag and SEBOV-GP; lane 3, Gag and ZEBOV-GP; lane 4, Gag and S33Z variant; lane 5, Gag and Z33S variant. (D) Supernatants from pcDNA3 (lanes 1)-, SEBOV-GP (lanes 2)-, and Z33S (lanes 3)-transfected cells were concentrated using Centricon Plus columns and analyzed as described for panel C. Panels C and D show the results of representative experiments, and similar results were obtained in two independent experiments.