Figure 1. Development of Nogo-A-specific siRNA.
(A-B) N2A cells grown on coverslips were transfected with siRNA-NS (A) or siRNA-NogoA (B) using TransIT-TKO transfection reagent (Mirus) for 48 hr. The cells were stained with goat anti-mouse Nogo-A (Santa Cruz) followed by Alexa Fluor 568-conjugated rabbit anti-goat IgG (red; Molecular Probes) and mounted onto slides with Vectashield containing DAPI (blue; Vector Labs). (C-H) The siRNA-NogoA was conjugated to fluorescein and injected intravenously into C57BL/6 mice with EAE. The spinal cords were removed 48 hr later, fixed, frozen, sectioned longitudinally and stained with DAPI. DAPI (C), flourescein-siRNA-NogoA (D) and merged images (E-G) illustrate cytoplasmic localization of the siRNA within cells of the lumbar region of spinal cords. (H) Sections were also stained with anti-APC to illustrate colocalization of siRNA (green) with oligodendrocytes (red) in the lesions.