Journal: bioRxiv

Article Title: Local aromatic interactions define temperature sensitivity of phase separation in an intrinsically disordered protein

doi: 10.64898/2026.05.07.723405

Figure Lengend Snippet: ( A ) Temperature-dependent droplet formation of Sup35 NM, N, and M domains monitored by turbidity at 500 nm in the presence of 15% (w/v) PEG 20,000. ( B ) Representative optical microscopy images showing temperature-dependent changes in droplet abundance for Sup35NM, N, and M domains under 15% (w/v) PEG 20,000 conditions. Scale bars, 20 µm. ( C ) Schematic illustration of truncated Sup35NM variants: AM1NM, LSNM, and AM2NM. ( D ) Temperature-dependent turbidity profiles of AM1NM, LSNM, and AM2NM measured in 20% (w/v) PEG 20,000. ( E ) Representative optical microscopy images showing temperature-dependent changes in droplet abundance for AM1NM, LSNM, and AM2NM under 20% (w/v) PEG 20,000 conditions. Scale bars, 10 µm.

Article Snippet: The final reaction mixture was adjusted to contain 10 μM protein, 10 mM sodium phosphate buffer (pH 7.0), and 10–20% (w/v) polyethylene glycol (PEG, average molecular weight 20,000, Nacalai Tesque).

Techniques: Microscopy

Journal: bioRxiv

Article Title: Local aromatic interactions define temperature sensitivity of phase separation in an intrinsically disordered protein

doi: 10.64898/2026.05.07.723405

Figure Lengend Snippet: ( A ) Schematic representation of Tyr-substituted Sup35NM mutants (YA-NM, YAh-NM, YF-NM, and YFh-NM). Tyr positions are indicated by black lines, Ala substitutions by red lines, and Phe substitutions by green lines. ( B ) Temperature-dependent turbidity profiles of Tyr-to-Ala Sup35NM mutants (YA-NM and YAh-NM) in 20% (w/v) PEG 20,000. ( C ) Schematic representation of Tyr-substituted Sup35N mutants (YA-N, YAh-N, YF-N, and YFh-N). ( D ) Temperature-dependent turbidity profiles of Tyr-to-Ala Sup35N mutants, YA-N and YAh-N in 20% (w/v) PEG 20,000. ( E ) Blue-Native PAGE analysis of Sup35NM mutants carrying Tyr-to-Ala (red) or Tyr-to-Phe (green) substitutions. ( F ) Temperature-dependent turbidity profiles of Tyr-to-Phe Sup35NM mutants (YF-NM and YFh-NM) in 20% (w/v) PEG 20,000. ( G ) Temperature-dependent turbidity profiles of Tyr-to-Phe Sup35N mutants (YF-N and YFh-N) in 20% (w/v) PEG 20,000. ( H ) (left) Representative optical microscopy images showing droplet morphology of YF-NM and YF-N mutants at 20°C in 20% (w/v) PEG 20,000. Scale bars, 10 µm. Right panels show magnified views. Scale bars, 5 μm.

Article Snippet: The final reaction mixture was adjusted to contain 10 μM protein, 10 mM sodium phosphate buffer (pH 7.0), and 10–20% (w/v) polyethylene glycol (PEG, average molecular weight 20,000, Nacalai Tesque).

Techniques: Blue Native PAGE, Microscopy

Journal: bioRxiv

Article Title: Local aromatic interactions define temperature sensitivity of phase separation in an intrinsically disordered protein

doi: 10.64898/2026.05.07.723405

Figure Lengend Snippet: ( A ) ThT fluorescence images of YF-NM droplets formed in 20% (w/v) PEG 20,000 in the absence (top) or presence (bottom) of 2 M urea. Scale bars, 10 μm. ( B ) ThT fluorescence images of amyloid fibrils formed by YF-NM in the presence of 2 M urea. Scale bars, 10 μm. ( C ) Schematic representation of LS NM mutants (YAh- LS NM and YFh- LS NM). ( D ) Temperature-dependent turbidity profiles of WT-NM, YF-NM, and YA-NM in 20% (w/v) PEG 20,000 in the absence (black) or presence (blue) of 2 M urea.

Article Snippet: The final reaction mixture was adjusted to contain 10 μM protein, 10 mM sodium phosphate buffer (pH 7.0), and 10–20% (w/v) polyethylene glycol (PEG, average molecular weight 20,000, Nacalai Tesque).

Techniques: Fluorescence