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. 2019 Jan 15;294(10):3359–3366. doi: 10.1074/jbc.RA118.005103

Figure 2.

Figure 2.

A, BAA473 selectively induces secretion of IL-1β but not IL-6 and IL-8. PBMCs were primed with 0.1 ng/ml LPS overnight followed by BAA473 treatment for 16 h and HTRF detection of secreted levels of IL-1β, IL-6, and IL-8. Data presented are mean ± S.E. (error bars) (n = 3). *, p < 0.005 versus vehicle (paired t test). B–D, BAA473 induces secretion of cytokines as well as caspase-1 and LDH activity in THP-1 cells. THP-1 cells were primed with 1 μg/ml LPS overnight followed by compound treatment for 24 h. B, cytokine secretion in supernatant was measured by HTRF (*, p < 0.0007). C, caspase-1 activity was assessed using the Caspase-Glo® 1 assay. Nigericin was used as an NALP3 activator. D, LDH activity (*, p < 0.003) was measured using a colorimetric LDH assay kit. E, BAA473 induces cytokine production via ASC/CASP1. THP-1 cells primed with 1 μg/ml LPS were pretreated with 10 μm BAA473 or 5 mm ATP for 1 h, followed by overnight treatment with caspase-1 inhibitor (VX-765). The level of IL-1β secretion was measured by an HTRF assay. Data presented are mean ± S.E. (n = 3).