Cp110

Antibodies against acetylated α-tubulin, γ-tubulin, Flag (Sigma-Aldrich), acetylated lysine, α-tubulin, Cep164, ninein (Santa Cruz Biotechnology), TMEM67, MKS1, IFT140, IFT88, CP110 (Proteintech), HDAC6 (abgent), and GFP (Roche) were purchased from the indicated sources. Rhodamine- and fluorescein-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories, and Alexa Fluor 405-conjugated secondary antibodies were from Invitrogen. Horseradish peroxidase-conjugated secondary antibodies were from Santa Cruz Biotechnology. Tubastatin A and tubacin were obtained from Santa Cruz Biotechnology. DAPI and tetramethylrhodamine-conjugated phalloidin were from Sigma-Aldrich. Control and HDAC6 siRNAs and plasmids expressing HA-HDAC6, GFP-HDAC6, and GFP-α-tubulin were described previously38 (link)44 (link), and various mutants were generated by PCR and site directed mutagenesis. The plasmid expressing mCherry-β-tubulin was constructed by PCR using the pmCherry-C1 vector (Clontech). Plasmids expressing Flag-cortactin wild-type, 9KQ, and 9KR plasmids were obtained from Edward Seto (H. Lee Moffitt Cancer Center & Research Institute, Tampa, USA).

Cell lysates after various treatments were prepared and subjected to IB analysis as described (Gu et al., 2007 (link)). The sources of primary antibodies were as follows: Cullin-1 (Santa Cruz, CA), phospho-AKT-Ser473, phosphor-AKT-Thr308, pan-AKT, AKT1, AKT2, AKT3, Aurora A, VHL, Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11), PDK1, phospho-PDK1-Ser²⁴¹ and Rictor (53A2) (Cell Signaling, Denver, CO), γ-tubulin, α-tubulin Clone AA13, acetylated tubulin Clone 6-11B-1, β-actin (Sigma, St. Louis, MO), CP110, NDE1, Cep97, Arl13B (Proteintech, Wuhan, P.R.C). UBA3 (Abcam, Shanghai, P.R.C). EGFR and phospho-EGFR-Tyr845 were gifts from Dr. Wanhong Xu,

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.