Characterization of Atg18-Atg18 interactions with FRET based stopped-flow measurements. (A) Graph shows time courses of rapid mixing of 0.5 µM Atg18 C113 (labelled with Alexa Fluor 488) with 0.5 µM Atg18 C113 (labelled with Alexa Fluor 546) either in the presence of unlabeled liposomes (1 mM total lipid), Ins(1,3,5)P3 (6 µM) or free in solution. (B) Experiments are similar to (A), but Oregon Green and Texas Red were used as donor and acceptor dyes instead. (C) Measurements with Texas Red labelled liposomes in comparison with measurements using labelled proteins for monitoring of the FRET signal. Unlabelled protein was mixed with unlabelled protein bound liposomes to monitor aggregation by measuring of the absorbance at 405 nm. Time course traces were normalized for comparison.