Ab214427

Macrophages from each group were collected and lysed using RIPA lysis buffer, followed by protein quantification utilizing the bicinchoninic acid method. 10 μg total protein was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for separation and moved to the polyvinylidene difluoride (PVDF) membrane. 5% defatted milk powder in 0.5% TBST was prepared to block the membranes for 2 h. Primary antibodies against p-p65 (ab194926, 1: 1000, Abcam), p65 (ab16502, 1:2000, Abcam), SIRT1 (ab189494, 1:1000, Abcam), LOX-1 (ab214427, 1: 1000, Abcam), CD36 (GTX100642, 1:500, GeneTex, Shanghai) and GAPDH (GTX 100118, 1:10000; GeneTex) were employed to incubate the membrane at 4°C overnight. The membrane was rinsed 3 times by TBST and cultivated with Goat anti-rabbit secondary antibody (ab205718, 1:5000, Abcam) at room temperature for 2 h. The visualization and analysis of protein blots were carried out employing an ECL (Amersham) and Image J (version 1.52; National Institute of Health) [15 (link)].

After total protein was extracted from HUVECs using RIPA lysis buffer (Solarbio, China), the protein concentrations were measured using an Omni-Easy™ BCA Protein Assay Kit (ZJ102L; EpiZyme, Shanghai, China). Protein extracts were separated by SDS-PAGE, transferred to a 0.22-μm PVDF membrane, and incubated with primary antibodies against CHOP (A5462; Bimake), Bip (11587-1-AP; Proteintech, Wuhan, China), and LOX-1 (ab214427; Abcam, Cambridge, UK).