Characterization of hCdc14A-deficient cells. (A) Generation of a conditional-null hCdc14A cell line. (B) Southern blot analysis confirms biallelic mutations of the hCdc14A locus in hTERT-RPE cells. WT (+/+), flox, Δneo, and Δ alleles are marked. (C) RT-PCR analysis confirming expression of WT (arrowhead) and exon 2–deleted (asterisks) hCdc14A transcripts. (D) MI of asynchronous populations of Cdc14A^flox/+ and Cdc14A^Δ/Δneo cells. Cells were fixed and stained with Hoechst (n = 3; 300 cells per genotype). (E) Cdc14A^flox/+ and Cdc14A^Δ/Δneo cells were treated with Noco for 12 h followed by shake off into medium without Noco, fixation, and Hoechst (n = 3; 300 cells per time point). (F) Cells were fixed, stained for γ-tubulin, and categorized by centrosome number (n = 3; 200 cells per genotype). Error bars indicate mean ± SD.