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. 2021 Nov 1;32(21):ar34. doi: 10.1091/mbc.E21-07-0355

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© 2021 Sluysmans et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 1: — PLEKHA5 and PLEKHA6 interact with the N-terminus of PDZD11 through their tandem WW domains. (A) Schemes of human PLEKHA5, PLEKHA6, and PLEKHA7 showing amino acid positions and structural domains: WW (Trp-Trp) green, PH (pleckstrin homology) red, proline-rich (Pro-rich) blue, coiled-coil (CC) pink. Boxes show yeast two-hybrid (Y2H) screen preys. (B) Representative immunoblot (IB) analysis of PDZD11 in immunoprecipitates (IP) of either PLEKHA5 (P5) or PLEKHA6 (P6) from lysates of Caco-2 cells (preimmune serum as negative control) (n = 4 for P5, n = 3 for P6). (C) IB analysis, using anti-HA antibody, of GST pull downs of either PDZD11-HA (P11-HA) or CFP-HA (negative control) (preys, green), using as baits (red) either GST or GST fused to the indicated sequences. Bait degradation products are nonspecifically labeled in the CFP-HA pull down (red dashed rectangles). (D) IB analysis, using anti-GFP antibodies, of GST pull downs of GFP-tagged PLEKHA5 (full length), PLEKHA6 (full length), and PLEKHA7 (N-ter, 1-562) preys using as baits either GST or GST-PDZD11. (E) IB analysis, using anti-GFP antibodies, of GST pull downs of GFP-tagged PLEKHA5 (full length), PLEKHA6 (full length), and PLEKHA7 (N-ter, 1-562) preys (green) using as baits (red) either GST, or GST-PDZD11 full length, or the N-terminal 30 residues (P11-Nterm), or a mutant PDZD11 lacking the first 24 residues (P11-Δ24). Ponceau S-stained blots show baits (dashed black rectangles).