Figure 2.

HIPK2-depleted cells show aberrant midbodies. (A,B) Indicated HeLa cells were analyzed by IF 96 h post transfection at the indicated time post nocodazole (Noc) treatment on ice. Telophase cells, showing acetylated tubulin at the midbody, were scored. In (A), representative immunostainings are shown. Bar, 5 µM. In (B), the percentage of cells with acetylated tubulin at the midbody are reported as mean ± SEM from two independent experiments, each performed in duplicate, in which at least a total of 80 midbodies per condition were counted. (C–E) HeLa cells were transfected as in Figure 1A or with spastin-specific validated stealth siRNAs and analyzed 96 h post transfection by WB (C) and by IF (D,E). In (C), the arrow indicates the band corresponding to M87, the most abundant spastin form; M87∆4 was generally less abundant and barely detectable. In (D), representative immunostainings of siHIP2 and siSpastin telophase cells with aberrant midbody are shown. The arrows indicate tubulin-labeled puncta along the intercellular bridge. Bar, 5 µM. In (E), the percentage of cells showing aberrant midbodies is reported as mean ± SEM from three independent experiments, in which at least total 150 midbodies per condition were analyzed. ****p < 0.0001, χ² test.