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Figure 2


Fig. 2. IFT122A is a ciliary gene. (A) Northern analysis of IFT122A expression during cilia regeneration. CU428 wild-type cells were deciliated and allowed to regenerate cilia. Total RNAs extracted from log phase cells and from the cells collected at 0, 1 and 2 hours in the recovery buffer were probed with isotope-labeled IFT122A cDNA. The upper panel shows that IFT122A transcripts were highly induced during cilia regeneration in wild-type cells. The lower panel shows the ethidium bromide staining as loading control. (B) Detection of HA-tagged Ift122Ap in cilia by immunoblotting. Total proteins from cilia and cell body fractions were isolated from HA-tagged IFT122A, HA-tagged IFT172, and wild-type CU428 strains. In the upper panel, the blot was probed with anti-HA monoclonal antibody. A 140 kDa protein and 190 kDa protein were detected in the cilia fraction (Cil) from IFT122A-HA and IFT172-HA cells, respectively. In the lower panel, the same blot was re-probed with a control antibody against polyglycine. The cytoplasmic protein Pgp1p, shown as multiple high-molecular mass bands, was only present in the cell body fraction (CB).





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