Pcmv ha vector

The anti-TEX12 (ab122455), anti-pericentrin (ab28144), anti-gamma tubulin (ab191114), anti-centrin 1 and 2 (ab11257) and anti-SYCE2 (ab107745) antibodies were obtained from Abcam and anti-FLAG (F1804) antibody was purchased from Sigma. Human TEX12 sequences, corresponding to amino acids 1–123 (full length) and 49–123 (core) and 45–123, 1–113, 1–91, 1–56 and 87–123, and SYCE2 were cloned into the pCMV-HA vector (Addgene Catalogue no 631604) for mammalian expression with an N-terminal FLAG-tag.

The shRNA sequences of KTN1 were purchased from Sigma-Aldrich and its lentivirus plasmids, virus packaging, and cell infection were performed as described previously.^{38 (link)} Virus packaging and cell transfection were performed as described previously.^{12 (link)} The promoter of CXCL8 gene was generated and cloned into the pGL3-Basic vector (Promega). The full-length human NF-κB/p65 ORF was generated and cloned into the pCMV-HA vector (Addgene). To construct the parental plasmids for encoding human KTN1, the synthesized full-length coding sequence of human KTN1 was cloned into minicircle vector pMC.BESXP-CMV according to the previous report,^{39 (link)} the minicircle encoding human KTN1 was produced in the E. coli strain ZYCY10P3S2T22. The mutated CXCL8 promoter luciferase reporter plasmids were made by recombinant PCR for luciferase reporter assay. The cDNA target sequences of siRNAs (RiboBio CO., LTD) and/or shRNAs for KTN1, CXCL8, and NF-κB/p65 were listed in Table S3.