CD39 blockade renders macrophages hyperinflammatory. (A-C) Tnfα (A), Il-12/23p40 (B), and Il-10 (C) mRNA expression at 4 hours poststimulation in wild-type BMDMs stimulated with LPS in the absence (solid bars) or presence of 10 μM POM-1 (striped bars). The means ± SD of triplicate determinations are shown and are representative of 3 independent experiments. (D-F) Cytokine production by BMDMs stimulated with LPS in the absence (closed squares; solid line) or presence (open squares; dashed line) of 10 μM POM-1. TNF-α (A), IL-12/23p40 (B) and IL-10 (C) levels were detected by ELISA. The means of at least 3 independent experiments ± SEM are shown. (G-I) BMDMs were stimulated with LPS in the absence (closed squares; solid line) or presence (open squares; dashed line) of 10 μM POM-1 and then washed with phosphate-buffered saline (break in y-axis) and replaced with LPS-free media and monitored over the next 5 hours for TNFα (D), IL-12/23p40 (E), and IL-10 (F) production. Protein levels was measured by ELISA. The means ± SD of triplicates are shown and are representative of 3 independent experiments.