Immunohistochemical detection of AApoAII and catalase

We detected AApoAII deposition and catalase by immunohistochemistry (IHC) following a previously described method (Li et al., 2017 (link)). Antiserum against mouse ApoA-II was produced against guanidine hydrochloride-denatured AApoAII in our laboratory (Higuchi et al., 1983 (link)) and applied at a dilution ratio of 1:3000. Catalase antibody was applied (1:500, GTX110704, GeneTex Inc, CA, USA) to reveal the degree of peroxisome change in the liver. After incubation overnight at 4°C with the primary antibody, the sections were incubated with the biotinylated secondary antibody (1:300, DAKO, Glostrup, Denmark) for 1 hr at room temperature. Target proteins were identified by the horseradish peroxidase-labeled streptavidin-biotin method (1:300, DAKO). In the immunofluorescence experiments, the sections were incubated with the PPARα antibody (1:500, GTX101098, GeneTex Inc, CA, USA) overnight and incubated with Alexa Fluor 488 goat anti-rabbit antibody (1:500, Thermo Fisher Scientific, Japan) for 1 hr at room temperature and incubated with DAPI for 10 min. Images were captured immediately using a confocal laser fluorescence microscope (LSM 880 with Airyscan, Carl Zeiss, Germany). In a negative control section, the primary antibody was omitted to confirm the specificity of staining.