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. Author manuscript; available in PMC: 2022 Mar 15.
Published in final edited form as: J Immunol. 2021 Feb 10;206(6):1329–1336. doi: 10.4049/jimmunol.2000969

Figure 6. ASC release, oligomerization and phosphorylation is absent in ASC KO cells and restored by ASC wild type but not Y146 mutant knock in.

Figure 6.

Phosphorylated ASC in media was detected by 4G10 (A) and Y144 (B) antibodies after LPS+ATP stimulation of THP-1 cells for 30min+30min. After imaging, membranes were stripped and re-blotted with polyclonal ASC antibody. Arrows show ASC monomer (22 kDa), and dimer (44 kDa) and line shows oligomers (66 kDa and up). Cell extracts were also blotted for ASC to show relative levels of soluble ASC and actin in THP-1, WT ASC KI and Y146A ASC KI (C). The insoluble cell pellets of these cell extracts were analyzed for oligomerization as described (D). Images of YFP-ASC oligomerized specks (E) are compared to YFP-Y146A-ASC filaments (F). Scale bar is 125 μm. Experiments were repeated 4 times.