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. 2014 Mar 19;9(3):e91712. doi: 10.1371/journal.pone.0091712

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© 2014 Shi et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Immunoblot analysis of SEGFP and TNFR-Fc. After replacement by TNFR-Fc, SEGFP expression decreased in the sorted cell pools compared with that in each parental cell clone, but all sorted cell pools produced TNFR-Fc. (B) Outline of the genomic DNA analysis. Three primers were designed to analyze replacement of SEGFP. Integration of the GOI into a pre-selected site of SEGFP integration was demonstrated by band-shift. (C) Examination of the remaining (silenced) parental cassettes in SEGFP cell clones before (as a positive control) and after exchange (-S2 cell pools). SEGFP and HR vectors were used as controls for SEGFP cell clones and sorted cell pools, respectively.