Aortic tissues were immediately fixed for 2 hours in 4% methanol‐free formaldehyde. The fixed samples were then embedded into paraffin and were cut into slices. Heat‐mediated antigen retrieval was performed with citrate buffer. After blocking, paraffin‐embedded sections were stained overnight at 4°C with HMGB‐1 antibody and then with corresponding secondary antibodies for 1 hour. Next, 3,3′‐diaminobenzidine (DAB) was used as chromogen, and images were taken using a microscope.

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