Figure 1 from NLRP3 Inflammasome Activation Expands the Immunosuppressive Myeloid Stroma and Antagonizes the Therapeutic Benefit of STING Activation in Glioblastoma

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STING-induced NF-κB activation does not prime the NLRP3 inflammasome. A, THP1-Dual cells were incubated overnight with vehicle, MLRR (10 μg/mL), or LPS (1 μg/mL). The supernatant was harvested and NF-κB induction measured using the InvivoGen QUANTI-Blue secreted embryonic alkaline phosphatase detection system. B and C, BMDMs were incubated overnight with treatments as described in A. Cells were divided and processed for both IL-1β qPCR (B) and flow cytometry (C) as described in methods and stained intracellularly with eFluor780 αIL-1β proform to assess pro–IL-1β production. D, BMDMs were incubated overnight with priming drugs LPS or MLRR as described in A. Cells were then collected and treated for 2 hours in NLRP3 activator nigericin (10 µmol/L), inhibitor MCC950 (1 µmol/L), or combination as indicated. The supernatant was harvested and IL-1β secretion measured via R&D Systems murine IL-1β ELISA kit. Statistical significance was calculated using the Student t test. ns, not significant; ****, P < 0.0001. MFI, mean fluorescence intensity.

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