Fig. 2.

Nck2^−/− cells are multi-nucleated and display long protrusions. A, Mouse embryonic fibroblasts (MEFs) bearing Nck1 or Nck2 gene inactivation were analyzed by Western blotting to confirm NCK1/2 protein expression. B, Wild-type, Nck1^−/− and Nck2^−/− MEFs were analyzed by immunofluorescence for actin (red), tubulin (green) and DAPI (blue) to assess cellular morphology. Multi-nucleation (asterisks) and intercellular bridges (arrows) are indicated. Representative images are presented (scale bar = 40 μm). C, The penetrance of the multi-nucleation phenotype was calculated for each genotype. Mean values and standard deviation from three independent experiments with >100 cells each are presented (WT, n = 374; Nck1^−/−, n = 325; Nck2^−/−, n = 372) (****p ≤ 0.0001, Fisher's exact test).