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Axonal transport of recombinant baculovirus vectors

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1 Axonal transport of recombinant baculovirus vectors
Ying Li, Xu Wang, Haiyan Guo, Shu Wang  Molecular Therapy  Volume 10, Issue 6, Pages (December 2004) DOI: /j.ymthe Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

2 Fig. 1 Neuronal tropism of baculoviruses. (A to C) Internalization in PC12 neurons 0.5 (A), 2 (B), and 4 (C) h after adding Cy3-labeled viruses into the culture medium. (D to L) Confocal images of neuronal internalization of Cy3-labeled viruses in mixed primary cerebral cortex cell cultures (D to F), the rat striatum (G to I), and the retina (J to L). Many red viral signals are colocalized with green NeuN signals. GCL, ganglion cell layer; INL, inner nuclear layer. (M to O) Cy3-labeled viruses in the optical nerves. The green signals: NeuN immunostaining. The red signals: Cy3-labeled viruses. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

3 Fig. 2 Axonal transport of Cy3-labeled baculoviruses. Confocal images of colocalization of red viral signals and green NeuN or TH signals in (A to C) the lateral geniculate nucleus and (D to F) the primary visual cortex after intra-vitreous body injection and in (G to I) the cerebral cortex and (J to L) the substantia nigra after intrastriatum injection. (M) Virus genome detected by PCR in rat visual system after intra-vitreous body injection. Lane 1, 100-bp ladder; lanes 2 and 4, no injection and sham injection with 1× PBS as negative controls; lane 3, purified BV-CMV E/PDGF-luc as a positive control; lanes 5, 6, 7, 8, and 9, tissues from the retina, optic nerve, lateral geniculate nucleus, superior colliculus, and primary visual cortex, respectively. (N) Baculovirus genome detected by PCR in rat brain after intrastriatum injection. Lane 1, 100-bp ladder; lanes 2 and 4, no injection and sham injection with 1× PBS as negative controls; lane 3, purified BVCMV E/PDGF-luc as a positive control; lanes 5, 6, and 7, tissues from the cerebral cortex, substantia nigra, and striatum, respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

4 Fig. 3 Activity and specificity of the recombinant baculovirus vector BVCMVE/PDGF-luc. (A and B) Comparison of gene expression levels mediated by baculovirus vectors with three different promoters in primary neuronal cultures and primary glial cell cultures (A) 2 and (B) 6 days after infection. PDGF, human PDGF β-chain promoter; CMV E/PDGF, hybrid CMV enhancer/human PDGF β-chain promoter; CMV, human CMV enhancer/promoter. Results are expressed as RLU/mg protein + SD. (C to E) Neuronal specificity of BV-CMVE/PDGF-luc in the striatum. Red luciferase signals are colocalized with green NeuN signals. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

5 Fig. 4 Luciferase expression mediated by BV-CMVE/PDGF-luc. (A and B) Luciferase assay 2 days after intra-vitreous body (A) or intrastriatum (B) injection of BV-CMVE/PDGF-luc. Four rats were used for each experiment. Values are presented as means + SD. LGN, lateral geniculate nucleus; SC, superior colliculus. (C to E) Red luciferase signals are colocalized with green NeuN signals in the lateral geniculate nucleus. (F to H) Red luciferase signals are colocalized with green TH signals in the substantia nigra. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions


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