Fig. 3.

PTEN inhibition does not increase cell proliferation. DNA synthesis was followed by fluorescent microscopy detection of 5-ethynyl-2′-deoxyuridine (EdU) incorporation (red); the cell nuclei were counterstained with Hoechst (blue). Image analysis was used to quantify the percentage of proliferating cells (stained positive for EdU). A: subconfluent BEAS2B cultures were treated with increasing concentrations of bpV (phen) (nontreated control, 1 and 2 μM), and the samples were fixed and processed for detection of EdU incorporation after 24 h. Analysis of A1-A3 reveals a dose-dependent decrease in the number of proliferating cells during bpV (phen) treatment A4. B: a scratch wound was applied to confluent BEAS2B monolayers, and 24 h later the samples were fixed and processed for detection of the incorporated EdU; the initial wound area was recorded using bright-field microscopy immediately after wounding (yellow lines). Analysis of B1–B2 images reveals that bpV (phen) treatment induces a small decrease in the percentage of proliferating cells in the wound region (B3). Scale bars are 100 μm; data are means ± SE; n > 10; *P < 0.05.