Extended Data Fig. 4: LLPS of the mixtures of PSD-95 PSG with different SynGAP CC-PBM proteins at the physiological concentration.
From: O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95
a, Time-lapse fluorescence images showing LLPS of 5 μM PSD-95 PSG and recombinant SynGAP CC-PBM, semisynthetic SynGAP CC-PBM, SynGAP-S1159OG, or SynGAP-T1306OG, with 2% PEG8000 over 20 min. Scale bar, 10 μm. b, Box-and-whiskers plot shows statistical analysis of the liquid droplet areas of a. The horizontal lines mark the maximum, median and minimum values of the data, and boxes mark upper and lower quartiles. For each group, at least 30 fluorescence images from at least three independent experiments were analysed. Differences were assessed by one-way ANOVA followed by Tukey’s multiple comparisons test with 95% CI, p < 0.05 is considered significant. c, SDS-PAGE gel showing the distributions of PSD-95 and SynGAP proteins in the supernatant (S) and pellet (P) in the sedimentation-based assay. 5 μM PSD-95 or SynGAP proteins were mixed for 10 min at r.t. in the presence of the crowding reagent (2% PEG8000) and subjected with the sedimentation-based assays. Bar graph on the right shows quantification of the distributions. d, Bar graph shows quantification results in a. The quantification results are from three independent experiments and represented as mean ± SD. Differences were assessed by one-way ANOVA followed by Tukey’s multiple comparisons test with 95% CI, p < 0.05 is considered significant.
