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. Author manuscript; available in PMC: 2013 Mar 19.
Published in final edited form as: J Neurosci. 2012 Sep 19;32(38):13206–13220. doi: 10.1523/JNEUROSCI.5041-11.2012

Figure 1. Schematic illustrations of tetracycline-regulated (tet-off) lentiviral vector constructs and the experimental procedure.

Figure 1

(A, B) A sin lentiviral vector is used for the regulated expression of (A) NT-3 and (B) GFP. Regulated gene expression is driven by the CMV*-1 promoter. The coding sequence for the tetracycline transactivator tTa2s is driven by the constitutively active CAG promoter. Note that the NT-3 virus also contains a GFP expression cassette via an internal ribosome entry site (IRES) to allow for the detection of NT-3 producing cells. cPPT, HIV central poly-purine tract; WPRE, Woodchuck posttranscriptional response element. (C) Schematic illustration of the experimental procedure. 1, A wire knife (blue) was used to transect dorsal column sensory axons (red) at C2/3 level. 2, Immediately after dorsal column transections, BMSC (brown) were grafted into the lesion site. 3, Tetracycline-regulated (tet-off) lentiviral vectors expressing NT-3 or GFP (green) were injected into the midline of dorsal white matter 2.5 mm rostral to the lesion site. 4, Subsequently, rats underwent bilateral conditioning lesions of the sciatic nerve. 5, Three days before perfusion, CTB was injected into the sciatic nerves bilaterally to transganglionically label ascending sensory tracts (red).