Immunohistochemical Analysis of 8-OHdG

LV tissue sections were blocked with 10% horse serum in phosphate-buffered saline (PBS), incubated with 8-hydroxydeoxyguanosine (8-OHdG) antibody (MilliporeSigma, Burlington, MA) and then incubated with biotin-conjugated anti-goat IgG (Vector Laboratory, Burlingame, CA). The sections were then incubated with avidin and biotinylated horseradish peroxidase macromolecular complex (ABC, Vector Laboratory), and stained with 3-amino-9-ethylcarbazole (AEC) and hematoxylin (Vector Laboratory). The samples were examined under a light microscope (AX 10, Zeiss). The area and intensity of staining were blinded to score for quantification as we have described^{50 (link)}.