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Fig. 4 Biochemical Interaction between Seahorse and Dsh
Upper: Western blot with anti-Myc on lysates directly or samples pulled down with Ni-NTA (A and B) or anti-Myc (C). Lower: Western analysis with anti-Flag (Flg) (A and B) or anti-Seahorse (C).
(A) Flag and 6x His-tagged Seahorse (Flg-Sea-6H) was co-overexpressed with Myc-tagged GFP (Myc-GFP) or Dsh (Myc-Dsh).
(B) Myc-tagged Dsh (Myc-Dsh) were overexpressed in the presence or absence of overexpressed 6x His-tagged Seahorse.
(C) Myc-tagged GFP (Myc-GFP) or Dsh (Myc-Dsh) was overexpressed in the absence of overexpressed Seahorse. A faster migrating band could also been seen in the lower panel of pull-downs by anti-Myc. Since this band migrated faster than Seahorse and since it was present in both samples, it is likely to be nonspecific.
(D) Diagram of Dsh domain structure and different deletion and mutation constructs.
(E and F) Different versions of Myc-tagged Dsh were co-overexpressed with Flag and 6x His-tagged Seahorse (Flg-Sea-6H). Upper: Western blot with anti-Myc on lysates directly or samples pulled down with Ni-NTA. Lower: Western analysis with anti-Flag on lysates.
Reprinted from Developmental Cell, 14(6), Kishimoto, N., Cao, Y., Park, A., and Sun, Z., Cystic kidney gene seahorse regulates cilia-mediated processes and Wnt pathways, 954-961, Copyright (2008) with permission from Elsevier. Full text @ Dev. Cell