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. 2022 Aug 30;221(10):e202201050. doi: 10.1083/jcb.202201050

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© 2022 Dai et al.

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Figure 5. — ARL13 accumulates in BBSome-deficient cilia. (A) Western blot of isolated control (g1, WT) cilia treated with phosphatase inhibitor or protein phosphatase (PPase) as indicated and probed with antibodies against ARL13 and, as a loading control, IC2. (B) Western blot comparing full-length and regenerating control cilia; the latter were harvested ∼25 min after the onset of cilia regrowth. Note relative accumulation of IFT172 and PLD and the reduction of FAP12 and phospho-ARL13 in short growing cilia. The quantification of the band strengths normalized for those of IC2 is shown in brackets and is based on one experiment. (C) Western blot of control (g1, WT) and bbs4-1 cilia probed with antibodies against ARL13 and, as a loading control, IC2. (D) Western blot of pre-deciliation (pre) and regenerating bbs4-1 cilia (time in minutes after cilia amputation is indicated) probed with antibodies against ARL13, PLD, BBS4 and, as a loading control, IC2. Note the accumulation of ARL13 over time. The quantification of the band strengths normalized for those of IC2 is shown in brackets and is based on one experiment. This figure is also part of Fig. S3 B. See Fig. S5, B and D for analyses of other bbs mutants. (E) Western blot analysis of cilia from control (g1, WT) and fla10-1 maintained at the permissive temperature of 22°C or the restrictive temperature of 32°C for 2.5 h prior to cilia isolation. Three replica blots were probed with antibodies to IFT proteins, BBS4, ARL13, PLD, and as a loading control, IC2. Quantifications of the band is shown in brackets and is based on one experiment. (F) Model proposing that ARL13 is required for BBSome-PLD interaction. Source data are available for this figure: SourceData F5.