Fig. 9. LRS mutants modulate particle formation and LLPS of FL-N.

(A) The onset of particle formation observed by DLS. Z-average hydrodynamic radii as a function of temperature are shown for 3 μM ancestral FL-N (black), the LRS self-association enhancing mutant FL-N:G215C (magenta), and the abrogating mutants FL-N:L222P (cyan) and FL-N:L222P/R226P (blue), all in the absence (solid lines) and presence of 10 μM NA ligand T₁₀ (dashed lines). (B) LLPS was studied by optical microscopy using 5 μM samples of ancestral FL-N (left) or the LRS self-association suppressing mutant FL-N:L222P in the presence of 10 μM T₁₀ in 10.1 mM Na₂PO₄, 1.8 mM KH₂PO₄, 2.7 mM KCl, 10 mM NaCl, and 10% polyethylene glycol (pH 7.40) at room temperature. Images shown were taken after 6 hours.