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All experiments were performed at 72 h post transfection, for maximum transfection efficiency.
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HeLa-MTS hOGG1, HeLa-MTS-mutant hogg1, HeLa-Nuc- hOGG1, HeLa-Nuc-mutant hogg1 and HeLa-Vector were grown in 35 mm, 6-well culture plates for 72 h post transfection as this time point showed maximum transfection efficiency.
The nanoparticle encapsulation by microparticle formation was optimized to achieve lipoplex release and maximum transfection efficiency in surrounding cells.
Maximum transfection efficiency was recorded at optimum N/ P ratio of 3 : 1.
The induction frequency is much higher than that induced by transient transfection using the Amaxa Nucleofector, so far thought to achieve maximum transfection efficiency in DT40 cells [24].
pCMVβ-gal (Clontech) or pRL-CMV (Promega) was co-transfected to normalize for transfection efficiency.
The pRL-TK vector expressing Renilla luciferase was co-transfected to control for transfection efficiency.
In parallel, TK-Renilla was co-transfected as a control for transfection efficiency.
The nanocomplexes were further evaluated for their transfection efficiency in cells by screening the GFP signals with flow cytometry.
A FAM-labeled, random sequence pre-miR was used for monitoring transfection efficiency by flow cytometry.
All samples were corrected for the transfection efficiency with respect to protein concentration.
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