Development of steatotic mice and method of IRI. Five-week-old C57BL/6 male mice were fed either a normal diet (ND mice) or a high fat diet (HF mice) for 9 weeks (a). Blood supply to the cephalad lobes of the liver was interrupted for 60 min using an atraumatic clip. After 60 min of ischemia, the clip was removed. APC or saline solution was administered just prior to surgery and at 8 h and 16 h after reperfusion. Mice were sacrificed 4 h or 24 h after reperfusion (b). Representative H&E ((c)-(A), (c)-(B)) and Oil Red O staining ((c)-(C), (c)-(D)) of liver tissue after 9 weeks on the experimental diets. There was no steatosis in ND mice ((c)-(A), (c)-(C)). After 9 weeks of HF diet feeding, macrosteatosis was observed using H&E staining and Oil Red O staining ((c)-(B), (c)-(D)). The original magnification was ×100.