Traba, JavierAntón Hurtado, Olga MaríaAbdul-Sater, Ali2024-02-082024-02-082022Antón, O.M., Traba, J. (2022). Measurement of Cytosolic Mitochondrial DNA After NLRP3 Inflammasome Activation. In: Abdul-Sater, A.A. (eds) The Inflammasome. Methods in Molecular Biology, vol 2459. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2144-8_12978-1-0716-2143-1978-1-0716-2144-81064-374510.1007/978-1-0716-2144-8_12https://hdl.handle.net/20.500.14352/100473The NLRP3 inflammasome, a key component of the innate immune system that mediates caspase-1 activation, which in turn induces cleavage of the pyroptosis executioner gasdermin D and the proinflammatory cytokines IL-1β and IL-18, requires two signals to be activated. First, inflammasome priming is achieved after activation of Toll-like receptors, which leads to NF-κB signaling and transcriptional activation of the genes for NLRP3 and IL-1β. Next, the inflammasome complex is activated by a second signal that induces extrusion of mitochondrial DNA to the cytosol of the cell, which leads to its oligomerization by a not fully understood mechanism. Here we describe a simple method that employs quantitative polymerase chain reaction (qPCR) using SYBR green to measure the presence of mitochondrial DNA (mtDNA) in the cytosol, which can be used to measure cytosolic mtDNA levels after inflammasome activation.engbook part1940-6029https://doi.org/10.1007/978-1-0716-2144-8_12restricted access577.1InflammasomeQuantitative polymerase chain reactionMitochondrial DNANLRP3Biología molecular (Química)Bioquímica (Química)24 Ciencias de la Vida