Fig. 2. Silencing of FACE1 causes abnormalities in nuclear morphology. (A) Control HeLa cells (a) show a normal nuclear morphology whereas FACE1-silenced HeLa cells display a range of abnormal nuclear morphologies (b-h) when stained with Hoechst dye. The abnormalities included minor effects such as herniations in the nuclear periphery (b) and folds (c-f), or more severe effects such as lobules (f) and fragments (g) as well as formation of micronuclei (h). The number of cells with severe abnormalities increased significantly at later times after transfection. (B) Comparative statistics. Cells were divided into four classes. `Normal' nuclei have a morphology as in a; `folds and herniations' as in c-f, fragmentation as in g; and micronuclei as in h. The percentage of normal nuclei decreased from 60% at 24 hours to 20% at 72 hours. Folds and herniations reached a maximum (>40%) 48 hours after transfection and then decreased slowly. The number of cells with micronuclei increased throughout the observation time and reached a maximum (>30%) 120 hours post transfection. (C) Cells treated with FACE1 siRNAs in-phase superimposed with prelamin A stain (red) and Hoechst DANN stain (blue) to show micronuclei. Bars, 15 µm.