RNAi of cytohesin-1 strongly reduces in vivo migration of mature BM-DCs. Mature BM-DCs after transfection with control siRNA or cytohesin-1 siRNA (for silencing efficiency, see Western blot analysis of cytohesin-1 expression after RNAi of cytohesin-1, panel 1C) were labeled with 10 μM TAMRA or 1 μM CFSE, respectively. Cytohesin-1 knockdown DCs (2.5 × 105) and control DCs (2.5 × 105) were injected into the hind footpad of wild-type C57/BL6 mice. Flow cytometric analysis of cell suspensions from the extracted draining (popliteal) lymph nodes 24 to 30 hours after injection shows that in vivo migration of cytohesin-1 knockdown BM-DCs is strongly reduced, compared with control cells (A,B). To exclude fluorochrome effects on DC migration, the staining was switched between the experiments (C). Control BM-DCs are set to 100% migration (B,C). Error bars indicate ± SD. Each experiment was repeated 3 times independently.
