Morphological differences between wild type and TEF (−/−) fibroblasts.

<p>(A) The morphology of wild type (WT) and TEF (−/−) fibroblasts in culture was assessed by phase contrast microscopy. Scale bar: 40 µm. (B) Both cell populations were labeled red with phalloidin for staining the actin filaments and visualized by confocal microscopy. Scale bars: 20 µm. (C) Confluent cultures were mechanically disrupted, leaving an area devoid of cells, and fibroblasts were labeled green to determine the actin distribution at the leading edge. Scale bar: 20 µm. (D) The mRNA expression of different Rho GTPases was analyzed by quantitative RT-PCR. The expression levels in TEF (−/−) cells were referred to those in wild type cells. Histograms represent the mean ± SD of three independent experiments. (E) Distribution of cell cycle phases in wild type and TEF (−/−) fibroblasts was determined by flow cytometry after staining nuclei with propidium iodide. The percentage of cells in the different phases, G0/G1, S, and G2/M, is indicated.