Fig 3.

Cytolethal distending effect exhibited by A. actinomycetemcomitans OMVs on HeLa cells (A to E) and HGF (F to H). Cells were treated with strain D7SS OMVs isolated from plate-grown bacteria (wild type [WT]) for 24 h (A) and for 72 h (B and F), and with D7SS OMVs isolated from broth cultures (WT broth) for 72 h (D). As controls, cells were treated for 72 h with OMVs isolated from strain D7SS cdtABC cultivated on agar (cdtABC) for 72 h (C and G) and with PBS (buffer) (E and H). After incubation, actin filaments and nuclei were stained with phalloidin (green) and DAPI (blue), respectively. The left panels show merged images from staining with both dyes, and the right panels show images from DAPI staining only. Magnification, ×1,000. Bars = 10 μm.