ARL3 is undetectable at the TZ and basal bodies by immunostaining nor binds the IFT-B1-shed BBSome in cytoplasm. (A) arl3-282 ; ARL3 : HA : YFP-TG, arl3-282; ARL3Q70L:HA:YFP-TG, arl3-282; ARL3T30N:HA:YFP-TG, and arl3-282; ARL3ΔN15Q70L:HA:YFP-TG cells stained with α-CEP290 (red) and α-YFP (green). The CEP290-labeled TZ was shown. α-YFP fails to detect ARL3-HA-YFP and its variants. (B) Same cells as shown in A stained with α-IFT46 (red) and α-HA (green). IFT46-labeled basal bodies were shown. α-HA fails to detect ARL3-HA-YFP and its variants. (C) Immunoblots of α-YFP-captured proteins from cell body (CB) samples of HR-YFP (HA-YFP-expressing CC-125 cells), arl3-282; ARL3:HA:YFP-TG (in the presence of GTPγS or GDP), arl3-282; ARL3Q70L:HA:YFP-TG, and arl3-282; ARL3T30N:HA:YFP-TG cells probed for the IFT-B1 subunits IFT22 and IFT70 and the BBSome subunits BBS1, BBS4, BBS5, BBS7, and BBS8 in the absence of DTT (left) and in the presence of DTT (right). Input was quantified with α-YFP by immunoblotting. MW stands for molecular weight. ARL3-HA-YFP fails to recover the IFT-B1-shed BBSome in the cell body. For A and B, phase contrast (PC) images of cells were shown. Inset shows the proximal ciliary region and the basal bodies. Inset magnifications (100 times) were shown. Scale bars, 10 µm. Source data are available for this figure: SourceData FS5.