Th2 cytokine IL-4 has been previously shown to suppress the production of proinflammatory cytokines in monocytes and macrophages. However, the underlying molecular mechanism by which IL-4 signaling antagonizes proinflammatory responses is poorly characterized. In particular, whether IL-4 can modulate inflammasome signaling pathways is unknown. Here, we demonstrate that IL-4 suppressed NLRP3-dependent caspase-1 activation and the subsequent secretion of IL-1β but did not inhibit AIM2- or NLRC4-dependent caspase-1 activation in THP-1 and mouse bone marrow-derived macrophages. IL-4 markedly inhibited the assembly of the NLRP3 inflammasome, including ASC oligomerization, NLRP3-ASC interaction, and NLRP3 oligomerization. The negative regulation of the NLRP3 inflammasome by IL-4 was not due to the reduced mRNA production of proinflammatory cytokines. Supporting this observation, IL-4 did not inhibit LPS-stimulated NF-B signaling pathway. Furthermore, the IL-4-mediated suppression of NLRP3 inflammasome signaling was independent of STAT6-dependent transcription and mitochondrial ROS. Instead, IL-4 modulated subcellular redistribution of NLRP3 into mitochondria and microtubule polymerization upon NLRP3-activating stimulation. Our data collectively suggest that IL-4 could suppress NLRP3 inflammasome activation in a transcription-independent manner, thus providing an endogenous regulatory machinery to prevent excessive inflammasome activation.