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. 2020 Jan 10;9(1):178. doi: 10.3390/cells9010178

Figure 3.

Figure 3

IL-37 inhibits ASC oligomerization. (a) WT or IL-37tg macrophages were treated with LPS (500 ng/mL) or vehicle, as indicated. After 3 h, nigericin (10 μM) was added for another 2 h. ASC and β-actin were quantified. Depicted is one representative blot of three independent experiments. (b,c) ASC-cerulean reporter macrophages were treated with recIL-37 (100 pg/mL) for 1 h, as indicated, before being stimulated with nigericin (6 μM) or silica (250 ng/mL) for 90 min or 4 h, respectively. Control cells were treated with vehicle. Cells were imaged with confocal microscopy and images flattened to allow maximum-intensity projections of 3-dimenisonal deconvolved z-stacks. (b) The number of specks per field of view ± SEM is depicted. Results are from two independent experimental replicates, in each of which five fields per sample were imaged, and each field contained >100 cells. *, p < 0.05, **, p < 0.01, and ***, p < 0.001 for nigericin- or silica-treated cells (with or without recIL-37 pre-stimulation as indicated) compared to vehicle-treated cells that were not pre-stimulated with recIL-37. ###, p < 0.001 for recIL-37 compared to vehicle within the same stimulation condition (nigericin or silica). (c) Depicted are representative images of (b). White arrows indicate ASC specks; scale bars 20 μm. LPS: lipopolysaccharide, tg: transgenic, veh: vehicle, WT: wild-type.