Figure 3.

Centriole-associated CPAP is dynamic, and its loss does not destabilize already-assembled mother centrioles. (A) Experimental strategy used to explore the dynamics of mother centriole luminal CPAP in IAA-treated cells in B and C. Cells were treated with IAA to degrade CPAP and with centrinone to prevent centriole duplication. CPAP and γ-tubulin were immunolabeled, and centrosomes were imaged by wide-field and 2DSTED. (B) Centrosomes in IAA-treated samples have reduced or undetectable CPAP and normal γ-tubulin levels. (C) Quantification of CPAP signals. (D) Experimental strategy used to explore the dynamics of mother centriole luminal CPAP in CPAP siRNA–treated cells in E and F. (E) Centrosomes in CPAP siRNA–treated samples have reduced or undetectable CPAP and normal γ-tubulin levels. (F) Quantification of CPAP signals. (G) Experimental strategy used to explore long-term stability of mother centrioles after CPAP removal by IAA or siRNA used in H–J, Fig. 5 D, and Fig. S4 D. (H) Centrosomes in IAA or siRNA-treated samples have reduced or undetectable CPAP and normal γ-tubulin levels. (I) Expansion/2DSTED analysis of G1 centrioles after 60 h of IAA or siRNA treatment. Acetylated tubulin labels centriole MT walls. Acetylated tubulin is not superresolved. (J) Quantification of mother centriole phenotypes from expanded samples after 40 and 60 h of IAA and siRNA treatment. Scale bars: 0.5 µm (B, E, and H); 1 µm (I). n = centrosome number. *, P ≤ 0.05; ***.