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. 2010 Dec 9;5(12):e14185. doi: 10.1371/journal.pone.0014185

Figure 1. Establishment of GFP-tagged NRG1 transgenic mice with EF1α-promoter.

Figure 1

(A) A schematic illustration of a transgene construct carrying EF-1α genomic promoter, NRG1β1 cDNA, GFP-tag insertion, and poly A signal. (B) Estimation of the copy number of the transgene by PCR. The exon 3 fragment of NRG1 genome was amplified with 18–30 cycles using tail DNA from Tg5 and Tg7 and separated in an agarose-gel. (C) Protein lysate was prepared from whole brain of adult male NRG1-Tg mice (Tg5 and Tg7) and WT littermate and subjected to immunoblotting with anti-NRG1 and anti-GFP antibodies. A closed arrowhead marks the transgene products. (D) Quantification of mRNA levels for type-1 NRG1 by RT-PCR. cDNA fragments specific for type-1 NRG1 and GAPDH mRNAs were amplified in the presence of SYBR Green I. PCR amplification curves and difference in Ct were analyzed by a real-time temperature cycler (LightCycler, Roche Molecular Biochemicals). For figure display, RT-PCR products were also separated by agarose-electrophoresis and visualized with ethidium bromide staining.