Introduction: Systemic low- and high-grade inflammation in various acute and chronic diseases is associated with skeletal muscle mass atrophy and metabolic dysfunction. While the nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is an integral component of the innate immune system, its role in skeletal muscle atrophy and metabolic dysfunction is poorly understood. Complicating factors in understanding molecular mechanisms underlying this loss, are that muscle wasting is a multifactorial process, and human tissue under well-controlled wasting conditions is sparse. Purpose: Here we studied mechanistically how inflammation induces skeletal muscle atrophy and metabolic dysfunction. Methods: We treated differentiated C2C12 myotubes with different concentrations of lipopolysaccharide (0,10,100-200 ng/ml LPS), and measured muscle fiber diameter, gene expression and protein concentrations of markers of the NLRP3 inflammasome.  Results: LPS reduced fiber diameter up to 42±6% (at 100 ng/ml LPS) after 24 hrs, which remained smaller up to 72 hrs. NLRP3 and downstream caspase-1 mRNA gene expression increased in a dose-dependent response after 24 hrs. NLPR3 and cleaved (p20) caspase-1 effector protein concentrations increased dose- and time-dependently upon LPS treatment. The addition of 10 ng/ml TGF-β further induced NLRP3 gene expression upon LPS treatment. Using STED fluorescent imaging, NLRP3 colocalized to mitochondria (66±5%), with unknown functional consequences. Treating the C2C12 cells with additional SS31 (to stabilize cardiolipin function) mitigated LPS-induced fiber atrophy.  Conclusions: These data suggest an inflammation-induced priming of the NLRP3 inflammasome in myofibers, independent of immune cell involvement. We show a structural link between the NLRP3 inflammasome and mitochondria in skeletal muscle cells that requires further study to understand the functional role of the NLRP3 inflammasome in the development of skeletal muscle wasting and metabolic dysfunction.