1. Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates 
Simon Alberti, Amy Gladfelter, Tanja Mittag 
Cell 
Volume 176, Issue 3, Pages (January 2019)
DOI: /j.cell
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2. Figure 1 Schematic Phase Diagram
The coexistence line (black) separates the one-phase and two-phase regimes and is a function of environmental conditions such as temperature, pH, etc. The system does not undergo phase separation beyond the critical point.
(A) At concentrations below csat, the system is in the one-phase regime. At any condition within the two-phase regime, the system demixes into a light phase (with c = cL) and a dense phase (with c = cD). All conditions on a single tie line (the orange line is an example) result in two phase systems with fixed light-phase and dense-phase concentrations, cL and cD, respectively; only the volume fractions of the two phases, fL and fD, change relatively to each other (examples 2–4). The volume fractions resulting from demixing of condition 3 can be calculated by the lever rule and are fL = L ÷ T (i.e., the ratio of the lengths of L and T) for the light phase and fD = D ÷ T for the dense phase. In equilibrium, csat and cL are equivalent, but when phase separation is nucleated, csat and cL can differ during the ripening dynamics of the system.
(B) The spinodal (gray line) indicates the region of instability in which the system must undergo demixing via spinodal decomposition. In the area between the coexistence line, or binodal, and the spinodal, the system demixes when nucleated.
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3. Figure 2
Bioinformatic Analysis of the Amino Acid Sequence of FUS to Identify Protein Regions that Are Involved in Phase Separation
Schematic representation of the human FUS/TLS domain structure is shown on top. QGSY, region enriched for the residues glutamine (Q), glycine (G), serine (S) and tyrosine (Y) depicted in green; G-rich, region enriched for glycine residues depicted in blue; NES, nuclear export sequence depicted in magenta; RRM, RNA recognition motif depicted in yellow; RGG, region enriched for residues arginine and glycine depicted in orange; ZN, zinc finger domain depicted in purple; NLS, nuclear localization sequence depicted light blue; IUPred, prediction of intrinsic disorder; PLD, prediction of prion-like region (PLAAC); FOLD, intrinsic disorder prediction by PLAAC (black) and PAPA (purple). Fold index is shown in gray. Pi-Pi, pi interaction prediction; NCPR, net charge per residue (sliding window size of 10); FCR, fraction of charged residues; HYDRO, hydrophobicity (Kyte & Doolittle, sliding windows size of 9).
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4. Figure 3
Overview of Experimental Approaches Used to Evaluate Properties of Assemblies Formed by LLPS
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5. Figure 4 An Initial Functional Repertoire of Biomolecular Condensates
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