Fig. 1

PIP4KIIγ accumulates at the spindle pole and the distal ends of spindle fibers. a–c Representative images of the cellular distribution of PIP4KIIγ. a CGL2 cells were fixed and stained for PIP4KIIγ (green) with commercial antibodies form Sigma (a-i and ii) and from GeneTex (a-iii and iv) and α-tubulin (red). b, c Representative images of CGL2 cells stably or human fibroblasts (HFW) transiently expressing FLAG-tagged PIP4KIIγ (FLAG-2C) were fixed and stained for FLAG-2C (green) and α-tubulin (red). Nuclei or chromosomes were counterstained with DAPI (blue). Images were obtained using a fluorescence microscope (a, b) or an upright confocal microscope (c). Scale bar, 10 μm. d FLAG-tagged PIP4KIIγ associates with α-tubulin. Left, cells stably expressing empty vector (Neo) or FLAG-2C were subjected to immunoprecipitation (IP) using anti-FLAG antibodies. Right, cells stably expressing FLAG-2C were subjected to IP using mouse IgG or anti-α-tubulin (α-tub) antibodies. The immunoprecipitates were then analyzed by immunoblotting with the indicated antibodies