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. 2022 Mar 8;38(10):110469. doi: 10.1016/j.celrep.2022.110469

Figure 3.

Figure 3

Clustering of the UL112-113 IDR is sufficient for LLPS

(A) Binary classification of phase separation in vitro (from Figure S3B) as a function of UL112-113 (x axis) and KCl concentrations (y axis). White dots, no phase separation; green dots, phase separation.

(B) Representative images of UL112-113 droplets. LLPS was induced by mixing purified UL112-113 proteins with a low salt (75 mM KCl) buffer. Treatment with 500 mM KCl or 5% 1,6-HD was used to inhibit phase separation. 2 μg/μL of soluble mNeonGreen was mixed with a low salt buffer. Scale bar, 100 μm.

(C) Schematic illustrating the corelet system (adapted from [Bracha et al., 2018]). iLID-EGFP-ferritin forms a 24-mer corelet (green star). sspB-mCherry (red crescent), is conjugated to IDRs from different proteins. sspB fusion proteins bind to corelets upon blue light activation, leading to LLPS (blue).

(D) Schematic of constructs. mCherry-sspB was fused to the IDRs of UL112-113 p43, p84, HNRPA1c as a positive control, or left unfused as a negative control.

(E) Photoactivation of corelet-expressing cells as depicted in (D). (A–C and E) The data shown are representative of three independent experiments. Scale bar, 10 μm.