Artemisia Extract Suppresses NLRP3 and AIM2 Inflammasome Activation by Inhibition of ASC Phosphorylation

Figure 4

Inhibitory effect of APO on ASC oligomerization. LPS-primed BMDMs were pretreated with APO or KCl (150 mM), and ASC oligomerization in the cells was determined by Western blotting upon stimulation with (a) 5 mM ATP for 1 h, (b) 150 μg/mL silica for 3 h, (c) 10 μM nigericin for 1 h, (d) 2 μg/mL poly (dA:dT) for 1 h, and (e) 1.5 μg/mL flagellin for 3 h. β-Actin is used as an internal control. (f) LPS-primed BMDMs were stimulated with 5 mM ATP for 1 h, 150 μg/mL silica for 3 h, 10 μM nigericin for 1 h, 2 μg/mL poly (dA:dT) for 1 h, and 1.5 μg/mL flagellin for 3 h and stained with anti-ASC antibody for ASC specks (red) and by DAPI for nuclei (blue). ASC specks were marked with arrows, and (g) cells with ASC specks are presented as a percentage of the positive cells. Data were expressed as the mean ± SEM (). Statistical analysis was performed using Student’s t-test. and , compared with respective stimuli in LPS-primed BMDMs plus respective inhibitors. APO, A. princeps extract; BMDMs, bone marrow-derived macrophages.