FIGURE SUMMARY
Title

Evolutionary Proteomics Uncovers Ancient Associations of Cilia with Signaling Pathways

Authors
Sigg, M.A., Menchen, T., Lee, C., Johnson, J., Jungnickel, M.K., Choksi, S.P., Garcia, G., Busengdal, H., Dougherty, G.W., Pennekamp, P., Werner, C., Rentzsch, F., Florman, H.M., Krogan, N., Wallingford, J.B., Omran, H., Reiter, J.F.
Source
Full text @ Dev. Cell
Fig. 3

Defining an Evolutionarily Conserved Ciliome

(A) Venn diagram of the overlap of S. purpuratus, N. vectensis, and S. rosetta ciliomes. The white line encompasses the high-confidence ciliary proteins present in two or more ciliomes. Only proteins that possess a mouse homolog (BLAST E value ≤1e−5) are included.

(B) Immunofluorescent staining for cilia (ARL13B, red) and candidate proteins fused to GFP (green) expressed in IMCD3 cells. Human C4orf47-GFP localizes to cilia and cytoplasmic microtubules. Fusions of human CSNK1D and CCDC113 with GFP predominantly co-localize with the basal body component CEP164 (blue). For C4orf46-GFP, nuclei are stained with Hoechst (blue). Scale bar for whole cell images, 5 μm. Scale bar for cilia-only images, 2.5 μm.

(C) Ten GFP-tagged human proteins, out of 49 randomly selected proteins from the high-confidence ciliome, localize to cilia or the ciliary base of IMCD3 cells. Immunofluorescent staining marks GFP-tagged proteins (green), cilia are indicated by ARL13B or TUBac (red), as indicated, and the basal body is highlighted by CEP164 (blue). Scale bar, 2.5 μm.

(D) Immunofluorescent staining of human proteins fused to GFP (green) in the Danio rerio embryo (somite stage 6–10). Cilia are marked by staining for TUBac (red). Cilia from within the Kuppfer's vesicle and primary cilia found outside the Kuppfer's vesicle are depicted. Scale bar, 2.5 μm.

See also Figures S1 and S3; Tables S3, S4, and S5.
Fig. S1

Subcellular localization of select identified proteins, Related to Figure 3. (A) Immunofluorescence imaging of IMCD3 cell cilia (ARL13B or TUBac, red) and transfected fusions of GFP with randomly selected proteins from the human proteome. The GFP fusions localized to cilia or the ciliary base (green). The distal basal body is marked by CEP164 (white). Nuclei were stained with Hoescht (blue). (B) Immunofluorescence imaging of IMCD3 cells stained as in (A). GFP fusions of the human homologs of randomly selected high-confidence ciliary proteins. (C) Immunofluorescence imaging of three human homologs of randomly selected high-confidence ciliary proteins fused to GFP (green) in the D. rerio embryo (somite stage 6-10). Motile and primary cilia are labeled by TUBac (red) and DNA stained with Hoescht is shown in blue. All scale bars, 5 μm.

Acknowledgments
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Reprinted from Developmental Cell, 43, Sigg, M.A., Menchen, T., Lee, C., Johnson, J., Jungnickel, M.K., Choksi, S.P., Garcia, G., Busengdal, H., Dougherty, G.W., Pennekamp, P., Werner, C., Rentzsch, F., Florman, H.M., Krogan, N., Wallingford, J.B., Omran, H., Reiter, J.F., Evolutionary Proteomics Uncovers Ancient Associations of Cilia with Signaling Pathways, 744-762.e11, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell
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