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Transfer of Cre mRNA in blood chimeras.

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posted on 03.06.2014, 02:43 by Kirsten Ridder, Sascha Keller, Maria Dams, Anne-Kathleen Rupp, Jessica Schlaudraff, Domenico Del Turco, Julia Starmann, Jadranka Macas, Darja Karpova, Kavi Devraj, Candan Depboylu, Britta Landfried, Bernd Arnold, Karl H. Plate, Günter Höglinger, Holger Sültmann, Peter Altevogt, Stefan Momma

(A) Schematic drawings of the experimental strategies to test for recombination events in blood chimeras. Lethally irradiated ROSA26-LacZ mice receive BM cells from Vav-iCre-ROSA-GFP mice. The bone marrow of recipient mice was tested for engraftment by flow cytometry analysis of GFP expression (representative analysis in right panel; wild-type bone marrow, red line; Vav-iCre-ROSA-GFP donor bone marrow, green line; bone marrow of ROSA26-LacZ recipient mouse after engraftment, blue line). For adoptive transfer experiments, the same combination of transgenes was used with spleen and lymph node cells as donor organs. Representative flow cytometry analysis of GFP expression of donor (green line) compared to wild-type cells (red line). At the time of analysis, GFP-positive cells were undetectable in recipient spleens (blue line). Two months after bone marrow transplantation, recombined cells can be detected in the liver (B), granular cell layer (C), and Purkinje cell layer (D), as well as associated with blood vessels (E). None of the recombined LacZ-/X-Gal–positive cells were positive for GFP, excluding cell fusion. Scale bar, 10 µm (B–E).