Figure 6

Mutant bone marrow-derived cells in the recipient kidney do not express the epithelial marker pan cytokeratin. (A -F) FISH, confocal microscopy and three dimensional modeling was used determine the nature of Y chromosome staining (green) relative to the epithelial marker pan cytokeratin (red) and DAPI stained nuclei (blue). Panels on the left (A, C and E) are confocal images with XZ and YZ planes shown as side panels. The corresponding panels on the right (B, D and F) are 3D reconstructions of these confocal images. (A and B) A longitudinal-section of a tubule in a control male kidney showing dispersed Y chromosome signals (arrows) in cells with typical epithelial morphology and pan cytokeratin staining. (C- F) Y chromosome-positive mutant cells from the bone marrow (arrows) were found in close proximity to recipient pan cytokeratin positive epithelial cells, but failed to stain for this marker themselves. C and D are 4 wks after IR and E and F are 12 weeks after IR. Scale bars = 10 microns.