Hek293t cells rbc2202

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.

Antibodies, chemicals, and plasmids used in this study are listed in Supplemental Table S1. HEK293T cells (RBC2202; RIKEN BioResource Research Center) were cultured in DMEM with high glucose (Nacalai Tesque) supplemented with 5% fetal bovine serum (FBS). hTERT-RPE1 cells (CRL-4000; American Type Culture Collection) were grown in DMEM/Ham’s F-12 medium (Nacalai Tesque) supplemented with 10% FBS and 0.348% sodium bicarbonate at 37°C in 5% CO₂. IFT27-KO cells (cell line #IFT27-2-2) and CEP19-KO cells (cell lines #CEP19-1-2 and #CEP19-1-12) were established from hTERT-RPE1 cells, as described previously (Nishijima et al., 2017 (link); Zhou et al., 2022 (link)).