Extended Data Fig. 9: Dominant-negative effect of SynGAP T1306 O-GlcNAcylation on LLPS of PSD-95 PSG/SynGAP CC-PBM.
From: O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95
a, Schematic showing the procedures for forming the overall 25% O-GlcNAcylated SynGAP CC-PBM trimers with no, one, two, and three O-GlcNAc at the ratio of 42.2%:42.2%:14.0%:1.6%. b, Time-lapse fluorescence images showing LLPS of PSD-95 PSG with SynGAP CC-PBM at indicated concentrations and O-GlcNAcylated ratios over 10 min. Scale bar, 10 μm. c, SDS-PAGE gel showing the distributions of SynGAP CC-PBM and PSD-95 PSG in the supernatant (S) and pellet (P). 80 μM PSD-95 PSG were mixed with 80 μM partially O-GlcNAylated SynGAP CC-PBM with varied O-GlcNAcylation stoichiometry ranging from 10% to 100%. d, SDS-PAGE gel showing the distribution of SynGAP CC-PBM and PSD-95 PSG in the supernatant (S) and pellet (P). PSD-95 PSG at varied concentrations was mixed with SynGAP CC-PBM or 25% O-GlcNAcylated SynGAP CC-PBM at indicated concentrations. Unpaired two-tailed Student’s t-test with 95% CI was used to determine difference, p < 0.05 is considered significant. In c and d, bar graphs showing quantification of the distributions. The quantification results were represented as mean ± SD. In b-d, representative results are shown from three independent experiments.
