Figure 4. In-situ monitoring the evolution of interactions between KE and CMD. (A) Images of a selected region at different time points. (B) The time-dependent fluorescence co-localization analysis of the selected region. (C) The plot-line analysis of the selected region at 160 s. (D) The mixing of CMD-FAM and KE-RhoB resulted in an instant rising of emission at 575 nm, accompanied by a decline of emission at 512 nm, indicating the FRET effect between these two components. [CMD-FAM] = 0.2 mM, [KE-RhoB] = 0.05 mM. (E) As the assembly progressed, the emission at 575 nm declined while the emission at 512 nm gradually recovered, indicating the attenuation of FRET. (F) The evolutions of emission intensities at 512 nm and 575 nm.