NA14 and spastin localize to the centrosome and midbody during cytokinesis.

<p>(A) Merged images of endogenous NA14 (green) accumulated at the centrosome during interphase and at the midbodies during late cytokinesis, along with β-tubulin (red). (B) Endogenous spastin (green) localizes to the centrosome and the midbodies, as shown by co-staining for β-tubulin (red). Merged images are at the right. (C) HeLa cell lines stably expressing control shRNA (shCTL) or shRNAs against NA14 (sh3 shown) were immunostained for endogenous β-tubulin (green) and γ-tubulin (red), with merged images at the right. Bar: 10 µm. (D) Cell extracts from cell lines stably expressing the indicated shRNAs were immunoblotted for NA14. PLCγ1 (149 kDa) and β-tubulin levels were monitored to control for protein loading. # denotes a cross-reacting band. (E and F) Mitotic and multinucleated cells were quantified in control and NA14 shRNA stable cell lines (means ±SEM; <i>n</i> = 3, with 100 cells per experiment). Nuclei were identified by co-staining with DAPI. (G) Quantification of cell death in control and NA14 shRNA stable cells lines by measuring lactate dehydrogenase release from cells (means ±SEM; <i>n</i> = 3, with 100 cells per experiment). *<i>p</i><0.05; **<i>p</i><0.01.