Extended Data Fig. 4: Structure–function analysis of SYD-2 phase separation.
From: Assembly of synaptic active zones requires phase separation of scaffold molecules
a, Diagram showing the structure of SYD-2, including disordered regions and coiled-coils predicted by the indicated algorithms37,38,42. LH1/LH2, liprin-homology domains; SAM, sterile alpha motif domains. b, Purified recombinant mCherry–SYD-2 constructs tested in an in vitro phase-separation assay. mCherry was fused to the N terminus of each sequence. c, Left panels in each column, example time-lapse analyses of in vitro condensates comprising the indicated SYD-2 constructs, showing the presence or absence of liquid fusion behaviour. Right panels, FRAP dynamics of each SYD-2 condensate. Data are means ± 95% CI for at least three condensates from two independent experiments. The y-axes show percentage intensity. Phase-separation assays were performed in 20 mM Tris pH 7.4, 150 mM NaCl with 10 μM protein. FRAP assays were performed on condensates formed in crowding conditions of 10% PEG, except in the case of the blue curves, where there was no crowding agent. Green or blue curves indicate a positive result (scored LLPS+) and black curves a negative result (scored LLPS−). Scale bars, 5 μm.
