Ripa buffer

Total proteins were extracted using RIPA buffer (Euromedex, Souffelweyersheim, France), and then equal amounts of proteins were reduced, size-separated on 12% stain-free precast SDS-polyacrylamide gels (Bio-Rad, Hercules, CA, USA), and transferred to nitrocellulose membranes by using an iBlot2 apparatus (Thermo Fisher Scientific). The membranes were blocked in 5% milk in TBS-Tween 0.1% and incubated with specific primary antibodies overnight at 4°C; the antibodies were against ACE2 (AF933, R&D Systems, Minneapolis, MN, USA; 1:200), phospho- and total p38 (9211 and 9212, Cell Signaling Technology, Danvers, MA, USA; 1:2,000), phospho- and total NF-κB p65 (3039 and 8242, Cell Signaling Technology; 1:2,000), and β-actin (A2228, Sigma-Aldrich; 1:5,000). The blots were exposed to horseradish peroxidase-conjugated anti-rabbit (Cell Signaling Technology, 7074; 1:10,000) and anti-goat (A27104, Thermo Fisher Scientific; 1:2,000) secondary antibodies, and bound antibodies were detected using Clarity chemiluminescent substrate (Bio-Rad). Images were recorded using a Fujifilm LAS-3000 bioimaging system (Stamford, CT, USA).

DCA was from Santa Cruz Technologies. Galactose and glutamine were from GIBCO. Human anti-CD36-PE, anti-CD36L-PE, anti-LPR1-PE and IgG were from Beckton Dickinson and 7AAD from Beckman (Brea, CA, USA). The MEK5 inhibitor BIX02189 and the ERK5 inhibitor XMD8–92 were from Selleck Chemicals (Radnor, PA, USA). RIPA buffer to prepare protein extracts was from Euromedex (Souffelweyersheim, France). The complete protease inhibitor cocktail (Complete EDTA-free) and the phosphatase inhibitor cocktail (PhosSTOP) were from Roche (Bale, Switzerland). ERK5, ACDVL and MEF2A/C antibodies were from Cell Signaling Technology (Danvers, MA, USA) and Abcam (Cambridge, UK), respectively. The antibody against β-Actin and HRP-labeled secondary antibodies were from Sigma (Burlington, MA, USA).