Fig. 3

Blockade of THIK-1 suppresses ATP-evoked IL-1β release from microglia in human brain. A Comparison of expression levels (mRNA) of P2X7, THIK-1, ASC, NLRP3, Caspase 1 and IL-1β for major CNS cell types analysed by nuclear enriched transcript sort sequencing (NETSseq) of healthy human brain tissue donors (n = 32). B Workflow illustrating determination of IL-1β levels upon acute activation of microglia in human and murine brain slices by purinergic stimulation (5 mM ATP, 3 h) as a model to simulate acute brain pathology. C–E Effects on IL-1β release in response to THIK-1 activation by 100 µM ATP (C) or in addition by P2X7 activation with 5 mM ATP for 3 individual human donors (D), and in comparison to mouse brain slices (E). Note the requirement for high ATP (i.e., P2X7 activation) and the dispensability of a priming stimulus to trigger NLRP3-dependent IL-1β release in both human and mouse brain that is abolished by the NLRP3 antagonist MCC950. Data are normalized to control condition and are from four individual patients and four wild-type mice. P values refer to 5 mM ATP condition. F, G Suppression of IL-1β release triggered by 5 mM ATP upon blockade of THIK-1 and P2X7 in human (F) and murine brain slices (G). Data are normalised to 5 mM ATP condition and are from three individual patients or four wildtype mice. P-values refer to 5 mM ATP condition. Data information: data indicate mean ± SEM. Dots on bars show number of slices. P-values are from unpaired Student’s t tests