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Figure 7


Fig. 7. A heterologous cilia trafficking assay using transferrin receptor. LLC-PK1 cells stably over-expressing (A) wild-type transferrin receptor (hTFR), (B) (1-56)PKD2L1-hTFR and (C) (1-72)PC2-hTFR. All heterologous proteins have a triple HA epitope at their COOH-termini and are immunolabeled by anti-HA (green). Cilial axonemes are marked by anti-acetylated {alpha}-tubulin (red). (A) The wild-type transferrin receptor (hTFR) does not traffic into cilia. (B) Replacement of the cytosolic N-terminus of hTFR with the first 56 amino acids of PKD2L1 does not result in trafficking of the chimeric protein to cilia. The right-most panels in A and B are confocal sections through the cell body at the level of the nucleus to show the robust expression of the heterologous protein in all cells in the field. (C) Replacement of the cytosolic N-terminus of hTFR with the first 72 amino acids of PC2 results in expression of the chimeric protein in cilia, indicating this domain contains cilia-targeting information. Bars, 10 µm.





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