Figure 4 raw data

Raw experimental and computational data presented in Figure 4.Figure 4. GAP activity reveals bistability of the reconstituted network. (A) Effect of RabGAP-5 on Rab5 activation. Different concentrations of RabGAP-5 were present in experiments with 500 nM CF488A-Rab5:GDI and 2 μM GDI, before adding 80 nM Rabex5:Rabaptin5. Shown are time courses at increasing GAP concentrations. (B) Maximal rates kmax of Rab5 activation for curves shown in A. Without detectable activation within 150 min, the activation rate was set to 0 and the corresponding points are depicted in orange. Error bars are SD. (C) Activation delay Ti for data presented in A. Without detectable activation, the times to inflection point are denoted as >150 min (orange). (D) GAP addition after activation with 80 nM GEF. RabGAP-5 was added to 500 nM and 2 µM final concentration, respectively. The traces were offset to the point of GAP addition. (E) GAP titration response curve. The fold change was calculated by dividing the fluorescence intensity at steady state with the average fluorescence signal 10 min before GEF addition. +GAP → +GEF fold values (circles) are calculated based on traces in A and +GEF → +GAP values are taken from D. Activation is depicted in blue and inactive reactions in orange (fold