All inflammasomes require the adapter protein apoptosis associated speck-like protein containing a CARD (ASC) for the activation of caspase-1. After inflammasome activation, ASC assembles into a large protein complex, which is termed "speck". ASC specks can be observed as they reach a size of around 1 μm and in most cells only one speck forms upon inflammasome activation. Hence, ASC speck formation can be used as a simple upstream readout for inflammasome activation. Here, we describe a method for analyzing inflammasome activation by ASC speck visualization. First, we describe the generation of a clonal inflammasome reporter macrophage cell line overexpressing fluorescently tagged ASC. We then discuss stimulation conditions and the microscopic evaluation of ASC speck formation. © Springer Science+Business Media, New York 2013.
CITATION STYLE
Stutz, A., Horvath, G. L., Monks, B. G., & Latz, E. (2013). ASC speck formation as a readout for inflammasome activation. Methods in Molecular Biology, 1040, 91–101. https://doi.org/10.1007/978-1-62703-523-1_8
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