Figure 4.

KIF18A depletion causes spindle pole and chromosome congression defects in vitro. (A) Mouse spermatogonial GC-1 cells transfected with Kif18a or luciferase (Luc) siRNA for 24 h were fixed and stained with antibodies to KIF18A (green) and CREST (red). DNA was stained with 4′6′-diamidino-2-phenylindole (DAPI; blue). Representative results are shown. (B) Cells at various mitotic stages or with unaligned/misaligned chromosomes were summarized from GC-1 cells transfected with Kif18a or luciferase siRNA. The data were summarized from 3 independent experiments. (C) GC-1 cells transfected with Kif18a or luciferase siRNA for 24 h were fixed and stained with antibodies to α-tubulin (green) and γ-tubulin (red). DNA was stained with DAPI (blue). Representative images are shown. (D) GC-1 cells transfected with Kif18a or luciferase siRNA for 24 h were fixed and stained with antibodies to α-tubulin (green) and γ-tubulin (red). Mitotic cells with multiple spindle poles were summarized from 3 independent experiments. (E) HeLa cells transfected with KIF18A or luciferase siRNA for 24 h were examined under a light microscope. Representative images are shown. (F) Equal amounts of proteins from HeLa cells transfected with KIF18A or luciferase siRNA for 24 h were blotted for KIF18A and β-actin. (G) Mitotic cells of various stages or with unaligned/misaligned chromosomes were counted from HeLa cells transfected with KIF18A or luciferase siRNA for 24 h. The data were summarized from 3 independent experiments. (H) HeLa cells transfected with KIF18A or luciferase siRNA for 24 h were fixed and stained with antibodies to α-tubulin (green) and γ-tubulin (red). Mitotic cells with multiple spindle poles were summarized from 3 independent experiments.