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. 2015 Dec 10;44(6):e54. doi: 10.1093/nar/gkv1338

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — Levels of Neo^R expression from AAV-based targeting vectors carrying the IRES- or 2A-based promoter-trap systems. (A) Expression levels of the Neo^R mRNA (left) and protein (right). HCT116 and DLD-1 clones carrying a Hyg^R–5′ EGFP reporter were infected with EGFP-TV-IRES (IRES) or EGFP-TV-2A (2A). Parental HCT116 and DLD-1 cells were infected with AAV-based targeting vectors against PIGA exon 6 (PIGAex6-TV-IRES and PIGAex6-TV-2A) and intron 5 (PIGAint5-TV-IRES and PIGAint5-TV-2A). An AAV vector carrying no Neo^R gene was included as a control (VC) in each set of infections. The expression levels of the Neo^R mRNA and protein in the infected cells were determined by qRT-PCR (mean ± s.e.m.; n = 3) and western blotting, respectively, and shown after normalization to GAPDH. Two-tailed paired t-test was applied for statistical analyses of data. (B) Gel images of western blotting for the Neo^R and GAPDH proteins.