Figure 3 raw data

Raw experimental and computational data presented in Figure 3. Figure 3. Rab5:GDI activation is tuned by free Rab5:GDP abundance. (A) Rab5 cycles between the membrane and solution before and after nucleotide exchange. Top: sCy5-Rab5 molecule counts per frame and collective CF488A-Rab5 activation. Bottom: snapshots of the activation reaction. sCy5- and CF488A-Rab5 are depicted in yellow and cyan, respectively. The sCy5 channel was smoothed before merging to reduce non-specific high frequency noise. Scale bar is 10 µm. (B) Rab5 single molecule trajectories reveal GDP- and GTP-bound proteins on the membrane. Top: 500 tracks of membrane-bound sCy5-Rab5 particles before (GDP) and after (GTP) activation. Bottom: frequency diagram identifies two populations with distinct lifetimes. A monoexponential decay with lifetime τGDP and two-exponential decay with lifetimes τ1GTP and τ2GTP was fitted to grouped data from N = 5 independent experiments, respectively (nGDP = 4829, τGDP = 0.58 ± 0.04 s; nGTP = 3090, τ1GTP = 1.20 ± 0.35 s, τ2GTP = 10.6 ± 3.3 s; errors are 95 % CI). Boxplot with mean lifetimes, **P