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research article

Nanoscale organization of the endogenous ASC speck

Glueck, Ivo M.
•
Mathias, Grusha Primal
•
Strauss, Sebastian
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November 16, 2023
Iscience

The 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.

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1-s2.0-S2589004223024598-main.pdf

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openaccess

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CC BY

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7.03 MB

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f09e4fc77324a70d6d588edc7c78dcd1

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