Fig. 3.

The G2 centrioles in CEP295-depleted cells fail to recruit POC5 or POC1B and do not show post-translational modification of centriolar microtubules. (A,B) U2OS cells were treated with sicontrol or siCEP295 as shown in (A) and analyzed by immunofluorescence microscopy (B). Arrows in B indicate inhibited targeting of POC5 to nascent centrioles. (C–F) Rescue experiment. G-CEP295R-inducible cells were treated as described in (C) and analyzed by confocal fluorescence microscopy (D) or immunoblot analysis (E). (F) Histograms illustrating the percentages of cells exhibiting POC5 signals. Error bars represent the mean±s.e.m. from four independent experiments. The total number n as indicated. (G) PLK4–myc-inducible cells were treated with siCEP295 as described in Fig. 1C, analyzed by immunofluorescence staining with antibodies against: (i) POC5 and hSAS6; (ii) POC1B and hSAS6; (iii) hSAS6 and Ac-tub, and (iv) hSAS6 and Glu-tub, and expression in new-born centrioles quantified. Error bars represent the mean±s.e.m. n=3 independent experiments each scoring 100 cells. Scale bars: 0.5 μm in the enlarged figures.