Fig. 1

Priming of macrophages is essential to induce release of bioactive IL-1β. A and B PBMCs were primed with Pam₃CSK₄ and then stimulated with ATP, BzATP or nigericin for 3 h. Supernatants were analyzed (A) for IL-1β concentration by ELISA and (B) LDH-release. Results are expressed as % of maximal LDH-release. Mean + SEM (n = 3—4). C THP-1 monocytes were primed with Pam₃CSK₄ and afterwards stimulated with ATP, BzATP and nigericin for 6 h. IL-1β concentration in the supernatants was analyzed by ELISA. Mean + SEM (n = 3—5). D THP-1 macrophages were primed and stimulated as described in (A). IL-1β concentration in the supernatants was analyzed by ELISA. Mean + SEM (n = 3—4). Two-tailed two-sample t test, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. E THP-1 macrophages were primed with Pam₃CSK₄. Cells were then stimulated with ATP, BzATP and nigericin for 3 h. Supernatants from stimulated THP-1 macrophages were transferred on HEK-blue IL-1β reporter cells. SEAP production was detected by QUANTI-Blue and optical density was measured at 620 nm. Mean + SEM (n = 4). One-way ANOVA followed by Dunnett´s post-test, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001