FIG. 6.

Kinetics of Yersinia-directed Tir delivery and Y⁰ to Y′ modification. HeLa cells were infected at two MOIs—∼10:1 (bacteria/host cells) (A) and 100:1 (B)—for the time periods (minutes) indicated. Host cells were then fractionated by isolating both saponin-released cytoplasmic (Cyto.) and Triton X-100-soluble membrane (Mem) fractions. Samples were resolved by SDS-PAGE (6% polyacrylamide) for Western analysis probing with anti-HSV antibodies. (B) HeLa cells infected with Yersinia for 60 min at the higher MOI were treated with gentamicin for the indicated time (minutes), to kill the Yersinia, before fractionation. The positions of the Yersinia-delivered TirHSV Y⁰ and Y′ forms are indicated with arrows.