FIG. 3.

VipD is translocated into U937 cells in a Dot/Icm-dependent manner. U937 cells were incubated with the indicated strains at an MOI of 5 for 1 h, after which host cells were harvested, extracted with 0.2% saponin, and processed as described in Materials and Methods. SDS-PAGE-fractionated samples were immunoprobed with antibodies against VipD or ICDH. Wild type (WT): U937 cells incubated with Lp02. dotA: U937 cells incubated with Lp03. ΔvipD: U937 cells incubated with strain SV192.—: U937 cells with no bacteria added. WT-Bacteria: L. pneumophila Lp02 grown to postexponential phase and lysed in SDS sample buffer to detect VipD.