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本数据集包含图4中展示的原始实验与计算数据。图4：GAP（GTP酶激活蛋白，GTPase Activating Protein）活性揭示重构网络的双稳态特性。 (A) RabGAP-5对Rab5激活的调控作用。实验体系中预先加入500 nM CF488A-Rab5:GDI（鸟苷酸解离抑制剂，Guanine Nucleotide Dissociation Inhibitor, GDI）复合物与2 μM GDI，随后再添加80 nM Rabex5:Rabaptin5复合物，期间设置不同浓度的RabGAP-5。图中展示了GAP浓度递增条件下的时间进程曲线。 (B) 图A中各曲线对应的Rab5激活最大速率kmax。若在150分钟内未检测到激活，则将激活速率设为0，对应数据点以橙色标注。误差棒代表标准差（Standard Deviation, SD）。 (C) 图A中数据对应的Rab5激活延迟时间Ti。若未检测到激活，则将拐点到达时间记为>150分钟（橙色标注）。 (D) 先用80 nM GEF（鸟苷酸交换因子，Guanine Nucleotide Exchange Factor, GEF）激活后再添加GAP的实验。分别将RabGAP-5终浓度调整至500 nM与2 μM，所有轨迹均以GAP添加时刻为基准进行偏移对齐。 (E) GAP滴定响应曲线。倍数变化通过将稳态荧光强度除以GEF添加前10分钟的平均荧光信号计算得到。+GAP→+GEF组的倍数变化值（圆点）基于图A中的轨迹计算得出，+GEF→+GAP组的数值取自图D。激活反应以蓝色标注，未激活反应以橙色标注（fold