Anti cd36 antibody

For histological analysis, the samples were rinsed with PBS and the aortic roots were routinely embedded in OTC compound. The aortic roots were cut into 5 μm serial cryostat sections on the aortic valve plane. Cryosections were fixed in 4% paraformaldehyde for 30 min and rinsed in TBS. Nonspecific sites were blocked using the avidin/biotin blocking kit, followed by 1% BSA (Sigma) and 5% normal goat serum in PBS. Slides were incubated overnight at 4°C with anti-SR-A antibody (1:200, AbD Serotec), anti-CD36 antibody (1:50, Santa Cruz) and anti-CD68 antibody (1:100, Abcam) for macrophages. The slides were then rinsed and incubated with secondary antibody. The processed sections were visualised using an Olympus microscope (IX71; Olympus Corporation, Tokyo, Japan) and a fluorescent microscope (Olympus Microscope BX-51, Japan) or a confocal microscope (Alsi, Nikon). Means were taken from seven different mice, evaluating ten serial cryosections/tissue from each mouse. SR-A and CD36 were quantified by assessing the percent positive area of total plaque for each marker. Image-Pro Plus 6.703 software (Media Cybernetics) was used for the statistical analysis.

Cells were cultured in the confocal dish for 48–72 h and then washed with PBS for three times and fixed in 4% paraformaldehyde for 30 min, permeabilized with 0.5% Triton X-100, followed by blocking with goat serum, then cells were subjected to staining with anti-CD36 antibody (1:50, Santa Cruz), anti-β-catenin (1:100, Santa Cruz), anti-cmyc (1:100, GeneTex) or anti-GPC4 (1:100, Proteintech) antibody, followed by Alexa-488-conjugated goat anti-mouse secondary antibody (1:500, Bioss) or Alexa-647-conjugated goat anti-rabbit secondary antibody (1:500, Bioss) for imaging. The cells were then counterstained with 4′,6-diamidino-2-phenylindole (DAPI) as a nuclear indicator and imaged with a confocal laser-scanning microscope (Carl Zeiss, LSM 880 with Airyscan).