Figure 4.

Oxidised lipoproteins inhibit activation of NLRP3, noncanonical and AIM2 inflammasomes. (a) THP‐1 macrophages were subjected to oxidised HDL₃ or LDL (200 μg mL⁻¹) for 1 h and then to nigericin (4 μm) for 1 h. Cell culture media were analysed for IL‐1β concentrations by ELISA. Data are expressed relative to nigericin alone (control). (b) THP‐1 macrophages were subjected to oxLDL (200 μg mL⁻¹) for 1 h and then to SAA (3 µg mL⁻¹) for 5 h. The secreted mature IL‐1β (17 kDa) and caspase‐1 (20 kDa) were determined by Western blot from cell culture media. (c) The intracellular levels of pro‐IL‐1β (37 kDa), procaspase‐1 (45 kDa), ASC (22 kDa) and NLRP3 (108 kDa) were assessed from Western blots of the cell lysates. Data are expressed as band intensities relative to total protein loading per lane. (d) THP‐1 macrophages were subjected to LDL or oxLDL (200 μg mL⁻¹) for 1 h and then transfected with ultrapure LPS (2 µg mL⁻¹) for 6 h. Cell culture media were analysed for IL‐1β concentrations by ELISA (left panel) and for concentrations of LDH (right panel) by cell death assay. (e) THP‐1 macrophages were subjected to LDL or oxLDL (200 μg mL⁻¹) for 1 h and then transfected with poly(dA:dT) for 5 h. Cell culture media were analysed for IL‐1β concentrations by ELISA. The mean values shown are from 3 (a) and 4 (b–e) experiments, Western blots and the corresponding loading shown are representatives of 4 individual experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.