Mouse peritoneal macrophages were isolated according to a previous study13. After mice were euthanized, the outer layer of the peritoneum was incised using scissors, and 3% thioglycolate medium (2.5 mL) was injected into the peritoneal cavity. Then, the peritoneal cells were exuded from peritoneal cavities, washed and resuspended in DMEM/high glucose media (Hyclone, USA) with 10% fetal bovine serum (Gibco LifeTechnologies) and 1% (v/v) penicillin. Cells were then cultured in 6-well plates at a density of 1 × 106 per well at 37 °C. The nonadherent cells were gently removed 2 h after incubation, and the left pure macrophages were collected and cultured in 6-well plates. The primary culture of mouse peritoneal macrophages was then stimulated by Ang II and treated with SB225002 for further experiments.

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