Figure 3

Localization, uptake, and function of transplanted mitochondria. Representative immunofluorescent images (A) from ZDF rat hearts following direct injection of vehicle alone (Vehicle) or vehicle containing human mitochondria (Mitochondria) to the hearts of ZDF rats following 30 min of regional ischemia and 2 h, 3 days and 28 days recovery. Human mitochondria were used only to confirm mitochondrial uptake. The use of human mitochondria in a rodent model allows for the differentiation between native rodent mitochondria and transplanted human mitochondria based on immune reactivity to a monoclonal anti-human mitochondria antibody (MTC02). Human mitochondria are shown labelled with human mitochondrial MTCO2 antibody (MTCO2ab, red). Mito-Tracker Green FM was used to show the injected mitochondria and in situ mitochondria. Nuclei were stained using DAPI (blue). Merged images are shown on the far right. Images are shown enlarged and with specific areas for each image enlarged to show detail. Enlarged images are boxed in gold. Transplanted human mitochondria are indicated by arrows. Mitochondria were observed within and around cardiomyocytes. Scale bars are 15 um and 100 um in length. Total tissue ATP content in the area-at-risk mmol/mg protein in vehicle hearts and in hearts receiving diabetic ZDF or non-diabetic ZL-mitochondria at 2 h (B) and 28 days (C) recovery. All results are shown as the mean plus and minus the standard error of the mean for n = 6 for each group. Statistical differences are shown as *p < 0.05, **p < 0.01, ***p < 0.001.