Figure 3.

TPEN-induced IL-1β release is dependent upon caspase-1. (a) Lysates of mouse peritoneal macrophages treated with LPS (1 μg/ml, 2 h) and TPEN (10 μM, 4 h), or DTPA (1 mM) and pyrithione (50 μM), or TPEN±1 μM of 1-hydroxypyridine-2-thione (zinc salt) (ZnPyr), were blotted for the house keeping protein actin (band at 42 kDa) and for XIAP (band at 53 kDa) (i). Lysates were also blotted for active caspase-8 sub-units (43 and 18 kDa) (ii), and for pro- (45 kDa) and active (10 kDa) caspase-1 (iii). (b) LPS-primed primary peritoneal macrophages were treated with TPEN (10 μM, 4 h) plus and minus incubation with the caspase-1 inhibitor Ac-YVAD-CHO (100 μM) or the caspase-8 inhibitor IETD-CHO (100 μM) with cell death measured by LDH assay (i) and IL-1β release by ELISA (ii). All data are presented as the mean±S.D. from at least four separate experiments. Blots shown are representative. ***P<0.001