Supplemental References

homology domain 2; DAD, diaphanous autoregulatory domain. Lines represent the constructs used in this study. (B) FH2 nucleates actin assembly. Actin (2.5 µM, 10 % pyrenyl-labeled) was polymerized with the indicated concentrations of FH2 (in nM). (C) Same conditions as in (B), with FH1-FH2 as indicated. (D) Ionic strength dependence of the nucleating activity of mDia1. Actin (1.5 µM, 10 % pyrenyl-labeled) was polymerized in the presence of 100 nM FH2, and either 50 mM KCl (red curve) as used by Li and Higgs (2003), or 100 mM KCl (our standard conditions, blue curve). (E) Actin (1.5 µM, 5 % pyrenyl-labeled) was polymerized with BSA-coated beads (black curves) or with 7.6 pmol bead bound FH2 (red curves