Cxcl16

For immunohistochemical staining, 3 μm paraffin sections were placed in 10 mM sodium citrate buffer (pH=6.0) and heated in a microwave oven or pressure cooker for antigen unmasking after deparaffinization and rehydration. Then, the sections were blocked with 3% hydrogen peroxide and 5% bovine serum albumin. Next, the sections were reacted with primary antibodies against LDLr (1:200 dilution, Abcam, Cambridge, UK), HMGR (1:200 dilution, Bioss, Beijing, China), CD36 (1:400 dilution, Novus, Colorado, USA), CXCL16 (1:100 dilution, Bioss, Beijing, China ) and G protein coupled receptor 43 (GPR43) (1:200 dilution, Bioss, Beijing, China) overnight at 4 °C and subsequently incubated with biotin-labelled secondary antibodies. Finally, the sections were stained with diaminobenzidine and then discontinued in water when a brown colour was detected.
For immunofluorescence staining, cells were fixed and then treated with 0.02% Triton X-100 for 15 min followed by blocking with 5% BSA for 1 h. Next, the cells were incubated with primary antibodies against LDLr, HMGR, CD36, CXCL16 and GPR43 overnight at 4 °C, followed by treatment with secondary antibodies conjugated to Alexa Fluor 555 for 1 h. Nuclei were stained with 4,6-diamidino-2-phenylindole. Finally, cells were examined by Olympus fluorescence microscopy.