Cep63 and Cep152 Cooperate to Ensure Centriole Duplication
Figure 3
Cep63 is needed for efficient centriole duplication in human and mouse cells.
(A-B) Cep63 depletion by RNA interference (RNAi) led to decreased centriole numbers in U2OS cells. Cells treated with control or Cep63 (Cep63-2) short interfering RNAs (siRNAs) for 4 days were stained with anti-centrin 2 (red) and anti-Cep63 (green) antibodies, plus DAPI (blue) to visualise DNA. Small panels show 3 times enlargements of the centrosome region with centrin 2 staining. Scale bar is 5 µm. (B) The number of centrin foci per mitotic cell was scored in three independent experiments, n>60. (C) Cep63 homozygous gene-trap mouse embryonic fibroblasts (MEFs) are devoid of Cep63 protein. Primary MEFs, at passage 3, were stained with anti-Cep63 (green) and anti-γ-tubulin antibodies (red), and DAPI (blue) to detect centrosomes and DNA, respectively. Representative images are shown of interphase and mitotic cells from two cell lines derived from littermates, one homozygous wild type (Cep63+/+) and the other homozygous gene-trap (Cep63T/T). Scale bars 1 µm. (D-F) Cep63T/T MEFs have reduced centriole numbers. (D) Primary MEFs, passage 3, were stained with anti-centrin2 (green) and anti-γ-tubulin (red) antibodies and DAPI. Scale bars 1 µm. (E) Centrin foci were scored in mitotic cells from 3 different cell lines for each genotype at passage 3, 40<n<100. (F) Centrin foci were scored in all cells (all cell cycle stages) in passage 1 primary MEF cell lines from 3 Cep63+/+ and 3 Cep63T/T embryos, n >100.