Fig. 2: Phase-separated Nup98 FG domains show different behaviours at different scales.

a Recovery after bleaching of FG phases assembled from Mac98A FG domain or prf.GLFG_52 × 12 spiked with 2.5% Atto488-coupled FG domains (of the same species). Bleached area: manually defined to be circular with a diameter of ~3 μm. Fluorescence recovery over time is shown (recovery is normalized to 1 for a complete recovery). For prf.GLFG_52 × 12 the translational diffusion coefficient is shown, as derived from fitting the dataset to a theoretical exponential recovery curve (Methods). The Mac98A FG domain is essentially immobile. Although a slight increase of signal inside the bleached area was observed e.g., at 1800 s and 3600 s, this is likely due to self-recovery of the bleached fluorophore molecules but not due to diffusive recovery, as a change of postbleach profile⁹¹ is not observed. b 1D ¹³C spectra of FG phases assembled from Mac98A FG domain (top), prf.GLFG_52 × 12 (middle), and a thioflavin-T-positive, proline-free variant of prf.GLFG_52 × 12: (GGLFGGATNSQT)₅₂ (bottom), obtained by MAS NMR (at 36 °C, 20 kHz MAS). In red, the signal from direct excitation (showing all ¹³C atoms), in blue, the cross-polarization (CP) spectrum (selective towards rigid parts of the samples). All spectra were obtained by averaging 1024 scans.