Fig. 3.

DPF3 colocalizes with the chromosome passenger complex and bridging fibers during metaphase. (A) HeLa cells were co-stained for DPF3 (in green) and CENP-E, Aurora B, MKLP2 (also known as KIF20A) or PRC1 (in red). Merged channels and magnifications of boxed regions are shown. Images were captured with Zeiss LSM 880 Airyscan high-resolution microscope. Asterisks (*) indicate centrosomes. Scale bars: 10 µm. (B) Extracts from U2OS cells mock-transfected or transfected with FLAG–PLK1, HA–Aurora B or Myc–KIF2B plasmids were incubated with glutathione-conjugated sepharose beads conjugated with GST alone, GST–DPF3a or GST–DPF3b. Input and pulled-down proteins were detected by western blotting using the indicated antibodies. (C) U2OS cells were co-stained for DPF3 (in green or red) and KIF4A (in green), citron kinase (CT-K), MKLP2 and acetylated α-tubulin (in red). Individual channels, merged channels with DAPI nuclear staining (in blue) and magnifications of boxed regions are shown. Scale bars: 5 µm. Immunofluorescence and western blotting images are representative of two independent experiments.