Figure 6. shRNA-mediated knockdown of LRP6 expression inhibits Schwann cell differentiation within injured sciatic nerve.

(A) Images show morphology of GFP+ OPCs transduced in vitro with lentiviral-mediated control and LRP6-specific shRNA, scale bar 20 μm (B) FACS sorting strategy, numbers show percent of cells of a specified phenotype (C) Western blot analysis of LRP6 and tRFP protein level revealed efficient gene expression knockdown. (D) FACS-sorted GFP+ OPCs transduced with LRP6shRNA differentiate normally into MBP+ OLs, scale bar 20 μm (E) Differentiation of OPCs transduced with LRP6-specific shRNA into SCs is defective. t-Students test **p<0.01 (F) Confocal images show grafted GFP+/tRFP+ cells differentiated within host sciatic nerves at 4 weeks after implantation. Co-localization of GFP (in green), with MPZ or periaxin (in red) indicates SC differentiation of OPCs transduced with non-targeting control shRNA but not with LRP6 shRNA (scale bar 20 μm).