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. 2010 Dec 29;85(5):2060–2078. doi: 10.1128/JVI.02210-10

FIG. 2.

FIG. 2.

4070A virus-pseudotyping NSCs engraft and disseminate CasBrE Env vectors within the CNS. (A) Representative paraffin brain sections from mice transplanted with control, CasES, CasE, and hrGFP NSCs plus 4070A after immunostaining for β-galactosidase (β-gal; brown), a genetic marker engineered into C17.2 NSCs (53); total virus expression (Virus; brown); or CasBrE Env expression (Vector; brown/black). hrGFP vector expression was assessed in frozen brain sections by direct examination under epifluorescence illumination (lower right panel). Bar, 40 μm. (B) Semiquantitative immunoblot assessment of the 4070A virus/CasBrE Env protein expression levels from equivalent brain extracts of freshly frozen brain sections from mice transplanted with control, CasES, and CasE NSCs with or without 4070A. Samples were separated on 8% SDS-PAGE gels and immunoblotted with MAb 697 (top) and pig anti-AmLV antiserum (bottom). The results are representative of at least three separate mouse brains for each group and at least three separate samples per brain. (C) Virus titration analysis on 4070A-infected control, CasES, and CasE NSC-transplanted brains for total virus (white bars) and CasBrE Env-encoding virus (black bars). Virus titers were determined on samples taken from at least three separate transplanted brains in each group. Error bars indicate the standard deviations. All analyses were performed on mice sacrificed at 4 weeks after NSC transplantation.