Fig. 4

Depletion of PIP4KIIγ enhances MCAK accumulation at the spindle pole. a Representative images of cells immunostained for MCAK (green) and α-tubulin (red). The control or PIP4KIIγ-depleted CGL2 cells were fixed and stained for MCAK and α-tubulin. b Relative intensities of MCAK or α-tubulin at the spindle pole. Results are the mean ± SD of at least 200 mitotic cells from three independent experiments. *p < 0.05 and **p < 0.01 by Student’s t test as compared that in the control. c Depletion of MCAK by siRNA transfection. The control- or PIP4KIIγ-depleted CGL2 cells were transfected with 0, 25, or 100 nM MCAK-specific siRNA as described in “Methods” section. Twenty-eight hours after transfection, the cells were subjected to immunoblotting to examine the depletion efficiency of MCAK. d Depletion of MCAK prevents the formation of spindle defects in PIP4KIIγ-depleted cells. The percentage (mean ± SD) of mitotic cells containing abnormal mitotic spindles was determined using at least 500 mitotic cells from three independent experiments. **p < 0.01 by Student’s t test compared to cells that were not transfected with siRNA