Glueck, Ivo M.Mathias, Grusha PrimalStrauss, SebastianRat, VirgileGialdini, IreneEbert, Thomas SebastianStafford, CheAgam, GaneshManley, SulianaHornung, VeitJungmann, RalfSieben, ChristianLamb, Don C.2024-02-202024-02-202024-02-202023-11-1610.1016/j.isci.2023.108382https://infoscience.epfl.ch/handle/20.500.14299/204512WOS:001115069800001The NLRP3 inflammasome is a central component of the innate immune system. Its activation leads to for-mation of the ASC speck, a supramolecular assembly of the inflammasome adaptor protein ASC. Different models, based on ASC overexpression, have been proposed for the structure of the ASC speck. Using dual-color 3D super-resolution imaging (dSTORM and DNA-PAINT), we visualized the ASC speck structure following NLRP3 inflammasome activation using endogenous ASC expression. A complete structure was only obtainable by labeling with both anti-ASC antibodies and nanobodies. The complex varies in diameter between similar to 800 and 1000 nm, and is composed of a dense core with emerging filaments. Dual-color confocal fluorescence microscopy indicated that the ASC speck does not colocalize with the microtubule-organizing center at late time points after Nigericin stimulation. From super-resolution images of whole cells, the ASC specks were sorted into a pseudo-time sequence indicating that they become denser but not larger during formation.Nlrp3 Inflammasome ActivationRecruitment Domain ProteinNf-Kappa-BSuperresolution MicroscopyHuman MonocytesPyrin DomainAdapter AscCell-DeathBindingRedistributiontext::journal::journal article::research article