Figure 9: Intra-arterially transplanted cardiac endothelial cells engraft in the heart and other highly perfused tissues. GFP cardiac endothelial cells were stained with PKH-26 and transplanted intra-arterially via transthoracic injections. (a) PKH-26+ GFP+ cells can be found highly concentrated along the injection tract in the left heart ventricle. Note that PKH-26 fluorescence is brighter than GFP and thus this dual labeling method ensures more efficient screening for the localization of the donor cells in multiple tissues. (b) anti-GFP staining identifies donor cells in the injection tract and also migrating out of the tract forming vessels that are BS-1 lectin positive (pseudocolor in red). Arrows highlights the colocalization of GFP+ cells that are also BS1+ (yellow) migrating out of the injection tract. Scale bar represents 100 μm for both (a) and (b). (c) proteins from multiple tissues of four transplanted mice (1, 2, 3, 4) were processed and analyzed by western blotting for GFP. Positive control (heart from transgenic CAG-EGFP mice) shows a predominant band at 54 kDa which correspond to the GFP dimmer. Negative control is untransplanted heart.