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. 2018 Mar 5;217(3):1079–1095. doi: 10.1083/jcb.201610098

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© 2018 Beati et al.

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Figure 1. — Smash binds to Baz and Cno. (A) Domain structures of Baz and the Smash isoforms PM and PI. The region of Baz used as bait and the region of Smash isolated as prey in the yeast two-hybrid screen are indicated. Numbers correspond to amino acid residues in the respective proteins. (B) The PBM of Smash is recognized by the Baz PDZ2 and PDZ3 domains. Left: Overlay of a representative region of the ¹H–¹⁵N correlation spectra of the Baz PDZ1 domain in the absence (black) and presence of a 2-fold (blue), 6-fold (purple), and 12-fold (red) stoichiometric excess of the Smash PBM peptide. Middle and right: Same as left, except the Baz PDZ2 (middle) and PDZ3 (right) domain. (C) GFP-Smash PI binds to Baz in Drosophila embryos. Lysates of embryos expressing GFP-Smash PI were subjected to IP with anti-Baz (IP Baz) or the preimmune serum of the same animal as control (IP pre). Western blots were probed with the indicated antibodies. Bands corresponding to full-length GFP-Smash PI and Baz are indicated by asterisks. (D) Overlay of a representative region of the ¹H–¹⁵N correlation spectra of the Cno PDZ domain in the absence (black) and presence of a 2-fold (blue), 6-fold (purple), and 12-fold (red) stoichiometric excess of the Smash PBM peptide.