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. 2020 Jun 1;9(12):e015513. doi: 10.1161/JAHA.119.015513

Figure 3. Tranilast (TR) inhibits NLRP3 (nucleotide‐binding oligomerization domain, leucine‐rich repeat–containing receptor family pyrin domain‐containing 3) oligomerization and activation.

Figure 3

Low‐density lipoprotein receptor–deficient (Ldlr−/−) bone marrow–derived macrophages (BMDMs) (A through C), apolipoprotein E–deficient (ApoE−/−) BMDMs (D and E), wild‐type BMDMs (F and G), and J774A.1 macrophages (H through K) were treated as indicated. The supernatants (SNs), whole cell lysates (WCL) and the Triton‐insoluble fractions were analyzed by immunoblotting to determine the oligomerization of NLRP3 and apoptosis‐associated speck‐like protein containing a caspase recruitment domain (ASC), caspase‐1 activation, and the release of interleukin (IL)‐1β and ASC. L, Human embryonic kidney 293T cells were transfected as indicated. At 8 hours before harvest, cells were treated without (–) or with TR (50 μmol/L). The SN and WCL were analyzed by immunoblotting as indicated. DSS indicates disuccinimidyl suberate; F, Flag; HA‐NEK7, never in mitosis gene a‐related kinase 7; LPS, lipopolysaccharides; SDD‐AGE, semi‐denaturing detergent agarose gel electrophoresis.