cGAMP alone can serve as the priming signal by increasing mRNA levels for inflammasome components in a Sting- and Ifnar-dependent fashion. BMDMs were transfected with increasing amounts of 2′3′-cGAMP, dAdT, or addition of 200 ng/ml LPS and analyzed after 6 h. (a–f) qPCR transcript levels of inflammasome components Aim2, Nlrp3, Casp1, Il1b, and Asc in addition to Ifnb produced by BMDM after treatment. n = 3 independent experiments. Shown is a representative with technical replicates of three, mean ± SD. (g–j) IFN-β (g and h) and IL-1β (i and j) in supernatants from BMDMs treated with increasing amounts of 2′3′-cGAMP for 6 h (g and i) and 19 h (h and j). NT, not treated. WT, Ifnar^−/−, or Sting^gt/gt BMDMs were transfected with cGAMP, transfection reagent alone (TFXN), or not treated (NT) for 19 h. (k–m) IL-1β (k), IFN-β (l), or Tnf (m) levels of supernatants after treatment. (n–r) Transcript levels as measured by qPCR for inflammasome components Aim2, Nlrp3, Casp1, Il1b, and Asc produced by BMDMs after treatment. n = 3 independent experiments, mean ± SEM; ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.