Extended Data Figure 2 : Phase separation is an isoform-specific capability of phosphorylated HP1α that is perturbed by GFP fusion.

From: Liquid droplet formation by HP1α suggests a role for phase separation in heterochromatin

Extended Data Figure 2

a, 1 μl of a 400 μM solution of each protein was spotted on a plastic coverslip and imaged at 10×. Scale bars, 50 μm. Buffer was 75 mM KCl, 20 mM HEPES pH 7.2, 1 mM DTT. Phos-HP1α is phosphorylated in the N terminus and hinge, nE-HP1α has the N-terminal serines replaced with glutamate, Phos-GFP–HP1α is a N-terminal GFP fusion phosphorylated in the N terminus and hinge, Phos-HP1α–GFP is a C-terminal GFP fusion phosphorylated in the N terminus and hinge, Phos-HP1α(BPM) has the KRK hinge sequence mutated to alanines and is phosphorylated in the N terminus and hinge, Phos-HP1α–KCK has a C-terminal GSKCK tag added and is phosphorylated in the N terminus and hinge. b, Complete comparison of saturation concentration measurements between spin-down assay (left) and 340 nm turbidity-based measurement (right), some data are repeated from Fig. 1.