Nlrp3

Mice thoracic aortas were separated, optimal cutting temperature compound (OCT, CellPath, U.K.) embedded aorta frozen sections were prepared, and immunofluorescence staining was performed as described previously [23] . Frozen sections were fixed by 4% paraformaldehyde for 15 min, then permeabilized by 0.3% (v/v) Triton X-100 (Biosharp, Hefei, China) in PBS for 15 min and blocked with 5% (w/v) bovine serum albumin (BSA, Solarbio, Beijing, China) for 1 h, then incubated with primary antibodies against NLRP3 (1:100, Abways Technology, Inc., Shanghai, China), PYCARD (ASC, 1:100, Abways Technology, Inc., Shanghai, China), Caspase-1 p20 (1:100, Affinity Biosciences, OH, USA) and tert (1:100, Affinity Biosciences, OH, USA) at 4°C overnight, followed by Cy3-conjugated Goat Anti-Rabbit IgG (H + L) (1:100, Proteintech™, Wuhan, China) and Goat Anti-Rabbit IgG (H + L) Alexa Fluor 488 (1:100, Abways Technology, Inc., Shanghai, China) for 2 h, respectively. DAPI was used to stain the nucleus.
Treated VSMCs and ECs were fixed and blocked like the above procedures, and then incubated with γ-H2AX (1:100, Abways Technology, Inc., Shanghai, China), followed by Cy3, and then counterstained with DAPI.