Figure 2.

Depletion of vinexin reduces cell proliferation by delaying mitotic progression. (A) HeLa cells infected with a lentivirus expressing control (siCtrl) or shRNA against human vinexin (siVxn-1 or siVxn2) were harvested for immunoblotting assay of vinexin β and α-tubulin. (B) The proliferation rates of siCtrl and siVxn cells were monitored daily by Prestoblue assay. (C) Double thymidine-arrested siCtrl and siVxn cells were released to fresh medium for 12 h, then underwent α-tubulin immunostaining and Hoechst labeling. The midbody stage is when 2 daughter cells are still connected by the intercellular bridge, which is detected by α-tubulin immunostaining (arrows, a magnified example image is shown). Scale, 50 μm. The distribution of siCtrl or siVxn cells at the midbody stage and interphase expressed as mean ± SEM relative ratios with the ratio in siCtrl cells arbitrarily set to 1. Total cell number for each siRNA group from 3 independent experiments is > 1000. ** P < 0.01, * P < 0.05 by Student t test.