FIG. 4.

Isolation of DEN virus receptor complex from SK-SY-5Y and peripheral blood monocytes by pull-down assay. (A) Isolation of cell proteins with affinity to DEN virus E protein. Total proteins from human peripheral blood monocytes/macrophages (lane Ex) were incubated with NiNTA-agarose coupled with the recombinant E protein (lane R). Pulled down eluates (lane Er) were separated by SDS-10% PAGE and stained with Coomassie blue. Arrows indicate migration of the pulled-down eluted proteins. (B and C) Total proteins (lanes Ex) from U937 cells (B) and peripheral blood monocytes (C) were incubated with NiNTA-agarose alone (lane C) or coupled with the recombinant E protein (lane Er). Pulled down eluates were separated by SDS-10% PAGE, transferred to nitrocellulose membrane, and incubated with anti-HSP90 (upper panels) or with anti-HSP70 antibodies (lower panels). Western blots were developed by chemiluminescence. Molecular mass markers are indicated on the left.