Fig. 4

NMD-escaping mutations in dosage-sensitive genes cause abnormal phase separation. A AUC performance of predicting phase-separation-prone regions. B Schematic diagram of the wild-type and mutant SOX2 protein domains. C Phase diagram of wild-type SOX2 protein and NMD-escaping mutant SOX2 protein with 150 mM and 5 M NaCl concentrations. Scale bar, 10 µm. D Confocal images of live SOX2 knockdown HEK 293T cells transfected with mCherry tagged wild-type SOX2 protein, NMD-escaping mutant SOX2 protein and mCherry, and stained with Hoechst (blue). Scale bar, 5 µm. E Regression plot of mutation truncation percentage and TruncPS score of NMD-escaping mutations in haploinsufficient genes. F Left: schematic diagram of the wild-type and mutant PQBP1 protein domains. Right: model of the relationship between protein truncation length and phase separation ability. The longer the truncation length, the weaker the phase separation ability. G Phase diagram of PQBP1 NMD-escaping mutant proteins with 150 mM NaCl concentration. Scale bar, 20 µm. H Confocal images of live PQBP1 knockout HEK 293T cells transfected with mCherry tagged wild-type PQBP1 proteins, NMD-escaping mutant PQBP1 proteins and mCherry, and stained with Hoechst (blue). Scale bar, 5 µm