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. 2017 Sep 4;6(9):e006347. doi: 10.1161/JAHA.117.006347

Figure 7.

Figure 7

Trimethylamine‐N‐oxide (TMAO) induced vascular nucleotide‐binding oligomerization domain–like receptor family pyrin domain–containing 3 (NLRP3) inflammasome activation in a sirtuin‐3 (SIRT3)‐dependent manner in vivo. Eight‐week‐old female ApoE−/− mice were treated with or without 1% choline for 4 months. Mice were killed, and their aorta samples were collected immediately, snap‐frozen in liquid nitrogen, and stored at −80°C until required. A, Western blot detection of Ac‐SOD2, SOD2, and SIRT3 expression in aortas. B, Bar graphs showing quantification of the indicated proteins. Eight‐week‐old female wild type (WT) and SIRT3−/− mice were fed with or without 1% choline for 4 months. Mice were killed, and their aorta samples were collected immediately, snap‐frozen in liquid nitrogen, and stored at −80°C until required. C, Western blot analysis of caspase‐1 p20, NLRP3, Ac‐SOD2, SOD2, and SIRT3 contents in aortas. D, Bar graphs showing quantification of the indicated proteins. E, SOD2 enzymatic activity in aortas was assayed using a SOD1 and SOD2 Assay Kit with WST‐8 following the manufacturer's instructions. F, Measurement of caspase‐1 activity in aortas. G, Western blot analysis of IL‐1β, ICAM‐1, MMP‐9 and SIRT3 expression. H, Bar graphs showing quantification of the indicated proteins. Values are expressed as means±SE (n=10); *P<0.01 vs the vehicle‐treated control group; AU, arbitrary units; ICAM, intercellular adhesion molecule; IL, interleukin; MMP, matrix metallopeptidase; SOD, superoxide dismutase.