FIG. 8.

Entry of LcrV into the HeLa cell cytoplasm does not require pCD1-encoded proteins. Cytochalasin D-treated (5 μg/ml) HeLa cell monolayers were uninfected (A and B, column 1), infected with Y. pestis KIM8-3233.1 (lcrE yopM) (A, column 4) or KIM8-3233.1 expressing YopM from pTrcM.2 (B, column 4), and infected with pCD1⁻ Y. pestis KIM10 expressing HT-VN68 (A and B, column 2), LcrV (A, column 3), or YopM (B, column 3). IPTG was added to 0.1 mM to cultures for activation of inducible promoters. After 4 h, monolayers were fixed, permeabilized, and stained with α-HTV (A) or α-YopM (B). Proteins were detected by secondary staining with Oregon green-conjugated anti-rabbit IgG followed by visualization by confocal laser-scanning microscopy. A 63× objective was used; immunofluorescence (IF) and DIC images are shown.