Factors affecting NLRP3 aggregation: sample incubation time, protein concentration, temperature, and salt concentration.
1H-15N HSQC spectra of NLRP3 at pH 3.6. A, 200 μm protein concentration and 303 K (conditions used for structural analysis). Sharp monomeric signals are observed together with a central blurb coming from resonances of higher order species. B, same sample used in A after overnight incubation at 303 K. C, protein concentration of 650 μm, 303 K. D, protein concentration of 200 μm, 310 K. E, protein concentration of 200 μm, 303 K, and 100 mm NaCl.