Fig 4. Macrophage cytoskeleton remodeling is dependent on direct contact with live Eh.

(a) Eh were pre-treated for 5 min with 55mM D-galactose, or glucose as an osmotic control prior to incubation with macrophages for 5 min at a 20:1 ratio. After incubation, cells were washed, lysed and equal amounts of lysates were loaded onto SDS-PAGE gel and immunoblotted with the indicated antibodies. (b) THP-1 cells were stimulated for 5 min with live Eh at a 20:1 ratio, whole lysates of Eh, or with membrane or cytoplasmic fractions of an equal amount of Eh. Lysates were analyzed as in (a). (c) Increasing amounts of Eh secreted/excreted proteins (SP) were incubated with macrophages for 15 min. Equal amounts of lysates were loaded onto SDS-PAGE and analyzed by Western blot with anti-paxillin and anti-GAPDH.