Figure 3
Figure 3. ATP promotes regulatory macrophage development. (A-B) Tnfα (A) and Il-12/23p40 (B) mRNA expression in naïve wild-type BMDMs or BMDMs stimulated with LPS in the absence or presence of 100 μM ATP was determined after 4 hours of stimulation. The means of 3 independent experiments ± SEM are shown. (C) The percent increase of regulatory macrophage-associated transcripts in wild-type BMDMs in the presence of LPS and 100 μM ATP are shown at 4 hours poststimulation by qRT-PCR. The dashed line represents the expression of regulatory transcripts induced by LPS alone. The means of at least 3 independent experiments ± SEM are shown. (D-F) Secreted levels of TNF-α (D), IL-12/23p40 (E), and IL-10 (F) by wild-type BMDMs stimulated with LPS in the absence or presence of 100μM ATP at 8 hours poststimulation. The means of 5 independent experiments ± SEM are shown. (G) TNF-α (closed squares; black line), IL-12/23p40 (open squares; black line), and IL-10 (closed squares; gray line) secreted by wild-type BMDMs stimulated with LPS in the presence of increasing concentrations of ATP. Cytokine levels were determined after 8 hours by enzyme-linked immunosorbent assay (ELISA). The means ± SD of triplicate determinations are shown and are representative of 3 independent experiments. (H) The percent of TLR-induced cytokines produced in the presence of 100 μM ATP. Cytokine levels were determined after 8 hours by ELISA and are relative to TLR stimulation alone (dashed line). The means of TNF-α (black bars), IL-12/23p40 (striped bars), and IL-10 (gray bars) production ± SD of triplicate determinations are shown and are representative of 3 independent experiments. Statistical analysis is relative to samples stimulated with TLR stimuli alone. (I) LPS-induced TNF-α (black bars), IL-12/23p40 (striped bars), and IL-10 (gray bars) production by human macrophages exposed to 100 μM ATP alone or in the presence of 10 μM POM-1 was determined by ELISA at 8 hours poststimulation. The means ± SD of triplicate determinations are shown and are representative of 2 independent experiments.

ATP promotes regulatory macrophage development. (A-B) Tnfα (A) and Il-12/23p40 (B) mRNA expression in naïve wild-type BMDMs or BMDMs stimulated with LPS in the absence or presence of 100 μM ATP was determined after 4 hours of stimulation. The means of 3 independent experiments ± SEM are shown. (C) The percent increase of regulatory macrophage-associated transcripts in wild-type BMDMs in the presence of LPS and 100 μM ATP are shown at 4 hours poststimulation by qRT-PCR. The dashed line represents the expression of regulatory transcripts induced by LPS alone. The means of at least 3 independent experiments ± SEM are shown. (D-F) Secreted levels of TNF-α (D), IL-12/23p40 (E), and IL-10 (F) by wild-type BMDMs stimulated with LPS in the absence or presence of 100μM ATP at 8 hours poststimulation. The means of 5 independent experiments ± SEM are shown. (G) TNF-α (closed squares; black line), IL-12/23p40 (open squares; black line), and IL-10 (closed squares; gray line) secreted by wild-type BMDMs stimulated with LPS in the presence of increasing concentrations of ATP. Cytokine levels were determined after 8 hours by enzyme-linked immunosorbent assay (ELISA). The means ± SD of triplicate determinations are shown and are representative of 3 independent experiments. (H) The percent of TLR-induced cytokines produced in the presence of 100 μM ATP. Cytokine levels were determined after 8 hours by ELISA and are relative to TLR stimulation alone (dashed line). The means of TNF-α (black bars), IL-12/23p40 (striped bars), and IL-10 (gray bars) production ± SD of triplicate determinations are shown and are representative of 3 independent experiments. Statistical analysis is relative to samples stimulated with TLR stimuli alone. (I) LPS-induced TNF-α (black bars), IL-12/23p40 (striped bars), and IL-10 (gray bars) production by human macrophages exposed to 100 μM ATP alone or in the presence of 10 μM POM-1 was determined by ELISA at 8 hours poststimulation. The means ± SD of triplicate determinations are shown and are representative of 2 independent experiments.

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