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Fig. 1. FACS analysis of zebrafish fibroblast cells (Pac2) infected with serial
dilutions of the viral vector CLGY. Cells were infected with pseudotyped
virus. Two days post infection, cells were harvested and analyzed for YFP
expression by flow cytometry using a FACSCalibur Flow Cytometry Analyzer
(Becton Dickinson, USA). (A) Uninfected cells. (B) Cells infected with
concentrated CLGY stock (1.1x108 CFU/ml). (C) Cells infected
with 1:10 dilution of concentrated virus stock. (D) Cells infected with 1:100
dilution of concentrated virus stock. To calculate average number of provirus
integrations per cell, real-time quantitative PCR was performed on genomic DNA
isolated from infected cells. Genomic DNA from cells containing two copies of
an integrated provirus per cell was used as reference for provirus number,
while variations in template amount were normalized against an endogenous
target sequence (Chen et al.,
2002).
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