Figure 3
Figure 3. Production of cytokines by DCs matured with different TLR agonists with or without dasatinib. Immature DCs were stimulated with the respective TLR agonist: LPS (10 ng/mL), LPS derived from P gingivalis (1 µg/mL), Poly(I:C) (1 µg/mL) or R848 (2 µg/mL) combined with IFNγ (100 U/mL), or a cytokine cocktail consisting of IL1β (20 ng/mL), TNFα (20 ng/mL), and PgE2 (20 μg/mL). Either no dasatinib (dark gray) was added or dasatinib (50 nM) was added (light gray). At 16 hours later, supernatant was collected and was frozen for subsequent analysis. Analysis was performed using the FlowCytomix Bead Array (eBiosciences), following the instructions of the manufacturer. Data from 3 different experiments were analyzed. Values were transformed using the log of each measurement and were statistically evaluated by a paired t test. Data on cytokine production with similar stimulation conditions, but without IFNγ, are provided in supplemental Figure 2.

Production of cytokines by DCs matured with different TLR agonists with or without dasatinib. Immature DCs were stimulated with the respective TLR agonist: LPS (10 ng/mL), LPS derived from P gingivalis (1 µg/mL), Poly(I:C) (1 µg/mL) or R848 (2 µg/mL) combined with IFNγ (100 U/mL), or a cytokine cocktail consisting of IL1β (20 ng/mL), TNFα (20 ng/mL), and PgE2 (20 μg/mL). Either no dasatinib (dark gray) was added or dasatinib (50 nM) was added (light gray). At 16 hours later, supernatant was collected and was frozen for subsequent analysis. Analysis was performed using the FlowCytomix Bead Array (eBiosciences), following the instructions of the manufacturer. Data from 3 different experiments were analyzed. Values were transformed using the log of each measurement and were statistically evaluated by a paired t test. Data on cytokine production with similar stimulation conditions, but without IFNγ, are provided in supplemental Figure 2.

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