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Fig. 4
Harmonin and Cdh23 bind directly to each other, but not to Ift88. (A) Co-immunoprecipitation. MDCK cells were transfected as indicated by (+) with combinations of DNA vectors encoding Harmonin-HA, mbn-Cdh23-GFP, Cdh23-GFP or IFT88-HA. Input lanes: 1,4,7,10,13. Unbound fraction lanes: 2,5,8,11,14. Bound fraction lanes: 3,6,9,12,15. Immunoprecipitation was performed with an antibody against Harmonin (α-Harm) or GFP (α-GFP). (B,C) Confocal sections of transfected MDCK cells. (B) Double immunolabeling of mbn-Cdh23-GFP (green) and Harmonin (red). (C) Double immunolabeling of mbn-Cdh23-GFP (green) and Ift88 (red). Harmonin-HA: full-length zebrafish Harmonin isoform A fused to HA-tag at the C-terminal. mbn-Cdh23-GFP: zebrafish Cdh23 membrane bound cytoplasmic domain fused to GFP at the C-terminal. Cdh23-GFP: zebrafish Cdh23 cytoplasmic domain fused to GFP at the C-terminal. Ift88-HA: full-length zebrafish Ift88 fused to HA-tag at the C-terminal. Scale bar: 7.5 µm.