Fig. 3

P2X7-specific nbs influenced the P2X7 pathway in vitro. Primary microglia preincubated with P2X7-specific nbs showed a substantially lower calcium influx (monitored by Fluo4 [A]) and DAPI uptake (B) than those of the isotype control group after ATP (1.5 mM) challenge. As a positive control, ionomycin facilitated calcium influx across the plasma membrane. C In the absence of P2X7-specific nbs, stimulated microglia showed significantly higher caspase 1 activation (measured by the appearance of FAM-FLICA; n = 3) than ATP-activated microglia in the presence of these nbs. D Blockade of P2X7 by specific nbs dampened IL-1β release in cultured microglia after ATP and LPS stimulation. Both types of P2X7-specific nbs used in this investigation significantly decreased IL-1β release (n = 3). The findings in D are specified in E, showing that IL-1β release can be reduced by using even low doses of nbs. Statistical significance was determined by Student’s t test. ***p < 0.0001