Fig. 3

Influence of caspases, serine proteases, cysteine proteases and aspartic proteases on IL-1β release. A THP-1 macrophages were primed with Pam₃CSK₄ and then stimulated with ATP, BzATP or nigericin for 3 h. Inhibitors of Caspase-1 (Ac-YVAD-cmk) or pan-caspase (Z-VAD-fmk), serine protease (AEBSF), cysteine protease (E64), aspartic protease (pepstatin A) were added 1 h before stimulation. IL-1β concentration in the supernatants was analyzed by ELISA. ATP, BzATP or nigericin induced IL-1β release was set to 100%. Mean + SEM (n = 3—4). One-sample t test against 100%, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. B THP-1 macrophages were primed with Pam₃CSK₄ and then stimulated with BzATP in the absence or presence of KCl for 3 h. Pan-caspase inhibitor (Z-VAD-fmk) or serine protease inhibitor (AEBSF) was added 1 h before stimulation. IL-1β concentration in the supernatants was analyzed by ELISA. BzATP induced IL-1β release was set to 100%. Mean + SEM (n = 3—7). One-way ANOVA followed by Tukey´s post-test, ns ≥ 0.05 (C) THP-1 macrophages were primed with Pam₃CSK₄ and then stimulated with ATP, BzATP or nigericin for 3 h. Gene expression of PRTN3 and ELANE and CTSG was normalized to GAPDH. Mean + SEM (n = 3)