Htert rpe1 cells

Expression vectors for subunits of the IFT-A and IFT-B complexes and heterotrimeric kinesin-II used in this study are listed in Supplemental Table S1; most of them were constructed in our previous studies (Katoh et al., 2016 (link); Hirano et al., 2017 (link); Funabashi et al., 2018 (link)). The antibodies used in this study are listed in Supplemental Table S2. GST-tagged anti-GFP Nb (Katoh et al., 2015 (link)) and anti-mCherry Nb (the LaM-2 version) (Ishida et al., 2021 (link)) prebound to Glutathione Sepharose 4B beads (GE Healthcare) were prepared as described previously. SAG and Polyethylenimine Max were purchased from Enzo Life Sciences and Polysciences, respectively. HEK293T cells (RBC2202) and hTERT-RPE1 cells (CRL-4000) were obtained from the RIKEN BioResource Research Center and the American Type Culture Collection, respectively.

cDNAs for human CFAP20 (NM_013242.3) and FAM149B1 (NM_173348.2) were obtained from a cDNA library by PCR amplification. Human ICK cDNA was kindly provided by Takahisa Furukawa (Osaka University) (Chaya et al., 2014 (link)). Expression vectors for BROMI, CFAP20, FAM149B1, CCRK, and ICK and their deletion and point mutants and vectors for the production of lentiviruses expressing them are listed in Supplemental Table S1. Several of the vectors were constructed in our previous studies (Hamada et al., 2018 (link); Nakamura et al., 2020 (link); Noguchi et al., 2021 (link)). Antibodies used in this study are listed in Supplemental Table S2. GST-tagged anti-GFP Nb and anti-mChe Nb (the LaM-2 version) prebound to glutathione–Sepharose 4B beads were prepared as described previously (Katoh et al., 2015 (link); Katoh et al., 2018 (link); Ishida et al., 2021 (link)). SAG and Polyethylenimine Max were purchased from Enzo Life Sciences and Polysciences, respectively. HEK293T cells and hTERT-RPE1 cells were obtained from RIKEN BioResource Research Center (Catalogue No. RBC2202) and American Type Culture Collection (Catalogue No. CRL-4000), respectively.