Depletion of Cyclin A2 in NIH3T3 cells induces an accumulation of the cells in G2/M and a reorganization of Actin cytoskeleton and focal adhesion distribution. (A) Western blot analysis of Cyclin A2 depletion performed in noninfected, mock-infected cells (shLuc), or cells infected with a virus expressing an shRNA targeting Cyclin A2 (shCycA2). GAPDH is used as a loading control. Molecular markers are given in kilodaltons. (B) Cell cycle distribution of shLuc and shCycA2 NIH3T3 cells after release from a synchronization block. (C and E) Staining of F-Actin with phalloidin-rhodamine (red) and Vinculin (green) in NIH3T3 cells infected with shLuc or shCycA2 (C) or treated with the Cdk1 inhibitor RO 3306 or DMSO as a control (E). (insets) Fields with lower magnification. Bars, 20 µm. (D) FACS analysis of cell cycle distribution of NIH3T3 cells treated with RO 3306 (Cdk1 inhibitor) or DMSO as a control. Data are represented as means ± SEM (n = 3). PI, propidium iodide.