Extended Data Fig. 2: Phase separation assay investigations of resilin-like polypeptide in response to various biochemical conditions.
From: Programmable synthetic biomolecular condensates for cellular control
a, Salt-dependent and concentration-dependent formation of condensate based on different concentrations of RLPWT ([GRGDSPYS]20) (with 10% RLPWT-sfGFP) in 50 mM Tris with different concentrations of NaCl at pH 7.2, confirming its salt-sensitivity and the contribution of electrostatic interaction in condensate formation. Scale bar is 10 μm. n = 3 independent biological repeats with similar results. b, Evaluation of responsive phase behavior using 5 μM of RLPWT ([GRGDSPYS]20) (with 10% RLPWT-sfGFP) in 50 mM Tris and 2 M KCl at pH 7.2. Reentrant condensate formation at high-salt condition using 2 M KCl, indicating the contribution of hydrophobic residues to the condensate formation. Treating the condensate with hydrophobic molecule 1,6-hexanediol, which is known to disrupt non-ionic interactions in condensate, dissolves the condensate. In the meantime, treating the condensate with adenosine triphosphate (ATP), which is known to disrupt electrostatic interaction in the condensate, shows no effect on condensate integrity. The above results confirmed the contributions of non-ionic residues to condensate stability. Overall, the biochemical tests demonstrated that synIDPs can also be modulated by various biochemical cues just as its native orthologs. Scale bar is 10 μm. n = 3 independent biological repeats with similar results.
