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. 2009 Jul 27;77(10):4609–4620. doi: 10.1128/IAI.00301-09

FIG. 4.

FIG. 4.

PVs are decorated with the WT forms of EGFP-Cdc42 and -RhoA but not with EGFP-Rac1. (A) HeLa cells were infected as indicated in the legend to Fig. 1 and then transfected with pEGFP alone (a to c), pEGFP-Cdc42 WT (d to f), pEGFP-RhoA WT (g to i), or pEGFP-Rac1 WT (j to l) as described in Materials and Methods. After 24 h, cells were fixed and processed for immunofluorescence, using a specific anti-C. burnetii antibody (red) (b, e, h, and k). Cells were analyzed by confocal microscopy. Micrographs of representative cells are shown. PVs are indicated by arrows. Filipodia (d) and lamellipodia (g) are indicated by arrowheads. The colocalization between PVs and overexpressed proteins is shown in the merge column (c, f, i, and l). Bars, 5 μm. (B) Quantification of colocalization between PVs and EGFP fusion proteins. Results are expressed as means ± SE for at least three independent experiments. ***, P < 0.001. (C and D) Chloramphenicol treatment reduces the colocalization of PVs with EGFP-Cdc42 and EGFP-RhoA. Infected cells were transfected with pEGFP-Cdc42 WT (C) or pEGFP-RhoA WT (D) and then treated with chloramphenicol (Chl) as indicated in Materials and Methods. Colocalization between PVs and EGFP fusion proteins was quantified. Results are expressed as means ± SE for at least three independent experiments. *, P < 0.05; ***, P < 0.001.