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| Proteins, Expression, Isolation and Analysis | Lysates were centrifuged at 10,000 rpm for 10 min at 4C, and supernatants were incubated with anti-DYKDDDDK G1 Affinity beads (Genscript; L00432-10) for 3 h at 4C. Immunoprecipitation was performed using anti-DYKDDDDK G1 Affinity Resin (GenScript; L00432-10) for 3 h at 4C. After sonication and centrifugation, the cleared lysates were incubated anti-DYKDDDDK G1 Affinity Resin (GenScript; L00432-10) for 3 h at 4C. | Get A Quote |
Nuclear speckles (NSs) are viscoelastic network fluids formed via phase separation coupled to percolation (PSCP). Intermolecular crosslinks of SRRM2 lead to the emergence of system-spanning networks, although the physicochemical grammar governing SRRM2 PSCP remains poorly decoded. Here, we demonstrate that SRRM2 is extensively phosphorylated within the intrinsically disordered region (IDR), creating alternating charge blocks. We show that this specific charge pattern does not markedly alter the condensation threshold of SRRM2 in cells. Instead, SRRM2 charge blocks intensify intra-network molecular interactions to modulate the material properties of mesoscopic SRRM2 condensates. We further identify casein kinase... More
