The activated sensor recruits ASC through homophilic interactions of pyrin domains and ASC associates with pro-caspase-1 via CARDCARD interactions, a step needed to induce caspase-1 activation. The activation of caspase-1 results in the cleavage of IL-1b and IL-18 precursors to their mature forms and their eventual secretion. Several studies have found that the saturated FFA palmitate acid can trigger inflammation by activating inflammasomes. We tested whether the v3 FFA DHA might have the 4-Hydroxytamoxifen opposite effect on macrophages and suppress inflammasome activation, thereby reducing IL-1b secretion. The mechanism by which DHA increases autophagosome formation is unknown although a recent study indicated that mTOR controlled autophagy required intracellular calcium signaling. As both ATP and DHA are capable of eliciting an intracellular calcium flux we checked how the simultaneous addition of DHA and ATP affected the level of intracellular calcium in BMDMs. We exposed the cells to different concentrations of ATP, DHA, and to both signals together. We found that as previously described DHA triggered an increase in intracellular calcium. The addition of ATP alone elicited a higher increase in the peak intracellular calcium and a much sharper rate of increase that did DHA. The combination of ATP and DHA triggered a greater rise in intracellular calcium and a more prolonged increase than did either ATP or DHA alone. We are currently investigating whether the elevated intracellular calcium noted in these primary mouse macrophages is sufficient to trigger autophagosome formation. High concentrations of ATP have been shown to induced autophagy in human macrophages and macrophage cell lines. In the course of our studies, Yan et al. published a report demonstrating that v3 FFA suppressed macrophage NLRP3 and NLRP1b inflammasomes, but not AIM2 and NAIP5/NLRC4 inflammasomes. They found roles for FFAR4, FFAR1, and barrestin- 2 in v3 FFA signaling. They also demonstrated a ligand induced interaction between NLRP3 and b-arrestin-2. Our results differ slightly from those of Yan et al. We found a strong suppression of all the tested inflammasome activators, perhaps because we used a higher concentration of DHA and included the DHA in the I-AB-MECA priming step, thereby reducing NF-kB activation and limiting expression of some of the inflammasome components.