Fig. 5

Delayed cilium disassembly, cell cycle re-entry, and extended cell cycle length in mutant cells. a MEFs stained with antibodies against acetylated α-tubulin (green) and γ-tubulin (red). Scale bars: 2 μm. b Quantification of cilium length of WT (n = 80) and mutant (n = 67) MEFs serum starved for 24 h. c Quantification of cilium length of WT (n = 85) and mutant (n = 85) MEFs after serum re-addition for 24 h. d Quantification of the percentage of cells with cilia. MEFs were serum-starved for 24 h followed by re-stimulation with serum for 0, 12, or 24 h. e Representative imaging of MEFs with EdU staining (red). Hoechst stains nuclei. Scale bar: 100 μm. f Quantification of the percentage of EdU-positive cells out of total cells counted (~200 cells in each experiment). g Coronal sections of E16.5 cerebral cortex stained with EdU (red) and antibodies against CldU (green). Hoechst stains nuclei (blue). Right panels are enlargements of regions outlined by white dotted boxes in leftmost panels. Scale bar: 10 μm (leftmost panels); 20 μm (right panels). h, i Quantification of cell cycle length and S-phase length. j Representative imaging of E16.5 cortex sections stained with antibodies against Ki67 (red) and BrdU (green) after 24-h BrdU labeling. White arrowheads represent BrdU-positive and Ki67-negative cells. k Quantification of BrdU-postive and Ki67-negative cells out of total BrdU-positive cells. Error bars represent SEM of three independent experiments. *P < 0.05, ***P < 0.001, n.s represents not significant (Student’s t-test)