ASC speck formation as a readout for inflammasome activation
A Stutz, GL Horvath, BG Monks, E Latz - The Inflammasome: Methods and …, 2013 - Springer
A Stutz, GL Horvath, BG Monks, E Latz
The Inflammasome: Methods and Protocols, 2013•SpringerAll 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 …
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 …
Abstract
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.
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