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When appropriate, 100 ng of the expression vector for ERα/ERβ or for p53 family members (or a control empty vector) were included.
A shRNA targeting the luciferase gene (Clontech) and circularised empty vector were included as controls.
Control wells containing no plasmid DNA or empty vector were included for each transfection experiment.
Negative (no template) and positive controls (orf1 gene cloned in TA vector) were included in each PCR run.
COS-7 cells transfected with empty pXL-flag vector and Tm5NM-1(His)6 expression vector were included as a negative control.
For all transfections, 500 ng of the PB-CA-rtTA tet-promoter activation plasmid (Woltjen et al., 2009; Woltjen et al., 2011), and 580 ng of the appropriate pMF inversion-reporter vector were included in the DNA mix.
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Brief discussion on cellular uptake mechanism of non-viral vector is included to understand its utility for gene delivery.
In some experiments, 4 ng expression vector was included.
A pGL3-control-promoter (i.e. an SV40 promoter) vector was included in all assays.
This vector was included as a divergent immunogen to act as a likely negative control for the HA1-con vaccine, but also as a long-shot cross-reactive vaccine against many influenza viruses.
GST empty vector was included as a negative control.
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