IFNγ and cytosolic dsDNA regulate inflammatory caspase-5 and IL-1β release by epidermal keratinocytes.

A-B, Regulation of IL-1β and caspase-5 in Th1/Th17 cytokine-stimulated keratinocytes analyzed by RTqPCR and normalized to β-actin. Data represent mean + SEM, *, p < 0.05; **, p < 0.01; ***, p < 0.001, Student’s t test, n = 9. C, Keratinocytes stimulated with IFNγ, transfected with dsDNA [Poly(dA:dT)] ± DNase and the DNA-dependent IL-1β release was analyzed by ELISA. D, Representative immunoblotting of corresponding supernatants analyzed for DNA-dependent activation of caspase-5 (exposure times; caspase-5, 30s; cleaved caspase-5, 30min) compared to loading control (Ponceau staining), three independent experiments. E, Keratinocytes stimulated with IFNγ, transfected with dsDNA and indicated siRNA and the caspase-5 dependent IL-1β release was analyzed by ELISA. C, E, Data represent mean + SEM, **, p < 0.01; ***, p < 0.001, Student’s t test, n = 3–6.