Fig. 6: Kinetic model of sRSM-G4 coacervation.

a CD spectra of 10 μM G4 titrated with 0–40 μM sRSM. The CD signal readings (right) at two specific wavelengths, 198 nm and 265 nm, were utilized in the global numerical analysis. b SVD amplitudes (right) calculated from the dependence of 50 μM G4 imino NMR spectra (left) on increasing sRSM concentration. Asterisks mark removed baseline signal regions to improve accuracy in SVD analysis. c The optical density at 600 nm monitored during the titration of 10 μM G4 with 0–40 μM sRSM. n = 3 independent experiments; data are means ± s.d. d The initial phase of the reaction examined by stopped-flow fluorescence upon mixing 2.5 μM G4 with 0–20 μM sRSM. e Kinetics of condensation process monitored by light scattering (at 295 nm) upon rapid mixing of 2.5 μM sRSM with 0–20 μM G4 (left) or 2.5 μM G4 with 0–20 μM sRSM (right). Each trace (in d and e) represents the average of 3 or 4 measurements. Solid lines in graphs from (a) to (e) represent the best global fit to the kinetic data. f The minimal kinetic model describing the sRSM-G4 coacervation includes three main steps: (i) initial binding, (ii) formation of stable nuclei and (iii) growth leading to phase separation. The rate constants for the individual steps were derived from global numerical fit. g Free energy profile of sRSM-G4 assembly and the reported free energy values in (f) were calculated using the Eyring equation (298 K; 1 μM RSM; 1 μM sRSM-G4). h The simulation represents the dynamic equilibrium of individual species for 10 μM G4 and sRSM concentration ranging from 0 to 20 μM. i The concentration of soluble species monitored using spin-down assay after incubation of 10 μM G4 with varying concentration of sRSM. Solid line represents the simulated data of G4 in dilute phase (sum of G4, sRSM-G4 and (sRSM-G4)₂ species), obtained from a global numerical model; data points from 4 experiments. Source data are provided as a Source Data file.