Investigating Aldosterone Effects on Cell Types

The source and culture of RAW264.7 cells and fresh bone marrow cells (BMDMs) were the same as described in a previous paper [47 (link)]. HUVECs (Sciencell, Carlsbad, CA, USA) were cultured in endothelial cell medium (ECM, Sciencell, Carlsbad, CA, USA) containing 10% FBS, 1% endothelial cell growth supplement (ECGS), 1% penicillin and streptomycin, and maintained in a humidified atmosphere with 5% CO₂ at 37 °C. After the density of the HUVECs reached 80%, the cells were incubated in serum-free medium for 24 h prior to the cell experiments.
RAW264.7 cells, BMDMs, and HUVECs were divided into the CON group, ALD group (10⁻⁷ mol/L aldosterone was administered for 24 h), and ESA group (cells were pretreated with 10⁻⁶ mol/L esaxerenone 2 h prior to aldosterone treatment). In some experiments, HUVECs were induced with 50 ng/mL VEGFA (Proteintech, Wuhan, China, Cat#: HZ-1038) and treated with or without the VEGFA receptor blocker bevacizumab (4 μg/mL) (MCE, Shanghai, China, Cat#: HY-P9906), and HUVECs were treated with bevacizumab after aldosterone induction. HUVECs were induced with 10 ng/mL TGF-β1 (MCE, Shanghai, China, Cat#: HY-P70543) and treated with or without the TGF-β1 receptor blocker LY2109761 (2 × 10⁻⁶ mol/L) (MCE, Shanghai, China, Cat#: HY-12075), and HUVECs were treated with LY2109761 after aldosterone induction.