Figure 3.

Ezrin mediates TNF-α–induced inhibition of EC proliferation. (A–F) Overexpression of WT or DN ezrin does not affect actin cytoskeleton. ECs either untransfected (A) or transfected with WT or DN ezrin (C and E, respectively) and treated (B, D, and F) or not treated with 20 ng/ml of TNF-α were stained with rhodamine-phalloidin for the detection of actin filaments, using the protocol described elsewhere (59). (G) Quiescent BAECs transiently transfected with empty vector (EV), WT ezrin, or DN ezrin were cultured in serum (white bars) or serum containing 20 ng/ml of TNF-α (black bars) for 24 hours. Cells were labeled with BrdU (50 μg/ml) for the last 6 hours of culture. Cells were fixed and stained with anti-BrdU antibodies. BrdU-positive cells were counted in at least 6 different visual fields. Average data from 3 similar experiments are shown. *P < 0.01; **P < 0.001.