Fig. 2.

CDYL2 RNAi induces mitotic defects and genome instability. a IF staining of beta-tubulin and DAPI staining of Hela cells treated with siControl (i) or siCDYL2 (ii–vi) revealing micronuclei and DNA bridges between daughter nuclei ((ii), and a zoom of (ii) shown in (iii)), multinuclear cells (iv, v) and fragmented nuclei (vi). Arrows added to highlight certain defects. b Hela cells treated with control siRNA showed normal metaphase (i) organization of chromosomes and the spindle. However, those treated with siCDYL2 revealed a variety of abnormal spindle organization patterns as well as misaligned chromosomes (ii–v). c Mitotic spindle stability was assayed by cold-shock of Hela cells treated with either control (i) or CDYL2 (ii, iii) siRNA. Metaphase cells were enriched by pre-treatment with MG-132 for 2 h before cold-shock and fixation. IF was performed as in (a). d Selected still images from video microscopy analysis of Hela H2B-GFP cells treated with siControl (top) or siCDYL2 (lower). The times indicated correspond to the elapsed time after the first image shown in the time-lapse series. The experiment was repeated three times with similar results