Centrin 1

An anti-NudCL2 antibody was generated as described previously^{6 (link)}. Antibodies against centrin 1 (Millipore, Billerica, MA, USA), CP110 (Proteintech, Wuhan, China), γ-tubulin (Sigma-Aldrich, St. Louis, MO, USA), α-tubulin (Sigma-Aldrich, St. Louis, MO, USA), β-actin (Sigma-Aldrich, St. Louis, MO, USA), Plk1 (Sigma-Aldrich, St. Louis, MO, USA), Flag (Beyotime Biotechnology, Shanghai, China), c-Myc (Santa Cruz Biotechnology, CA, USA), SAS6 (Santa Cruz Biotechnology, CA, USA), separase (Santa Cruz Biotechnology, CA, USA), CDK2 (Santa Cruz Biotechnology, CA, USA), cyclin A (Santa Cruz Biotechnology, CA, USA), cyclin E (Santa Cruz Biotechnology, CA, USA), STIL (Abcam, Cambridge, MA, USA), HERC2 (BD Biosciences, San Jose, CA, USA; Bethyl Laboratories, Montgomery, TX, USA), Plk4 (Proteintech, Wuhan, China), and USP33 (Proteintech, Wuhan, China) were acquired commercially.

MDA-MB-435 and HOP-62 cells were incubated for 2 h on ice to induce the depolymerisation of cytoplasmic microtubules. Subsequently, cell lines were subjected to an IF assay using DAPI and two different marker combinations: (i) centrin-2 (1/500 dilution, Ref. sc-27793-R, Santa Cruz Biotechnology) and acetylated tubulin antibodies (1/500 dilution, Sigma, Ref. T7451), to visualise the long centrin-positive rod-like structures and stable MTs present in centriole and cilia, respectively; (ii) centrin-1 (1/1000 dilution, clone 20H5, Millipore) and ARL13B (1/500 dilution, kind gift from T. Caspary), to label the long centrin-positive rod-like structures and cilia, respectively (for more experimental details, Immunofluorescence staining section). The visual examination of the presence of acetylated tubulin, or ARL13B, on the long centrin-positive rod-like structures was performed on an Applied Precision DeltavisionCORE system (for representative pictures see Supplementary Fig. 4). These structures were considered as overly long when they were at least approximately twice the size of normal-length centrioles. Three independent experiments were performed, with at least 30 overly long centrioles analysed per condition and per experiment.