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. 2014 Nov 11;42(21):13061–13073. doi: 10.1093/nar/gku1124

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© The Author(s) 2014. 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. — Competition between tTA and tetTALE/tetTALE-SD. (A) Experimental setup. X1/5 cells containing a P_tet-luciferase reporter and a tTA expression cassettes stably integrated were cultured under ON conditions (-dox) where tTA is bound to tetO. Cells were stably transfected with either tetTALE or tetTALE-SD expression construct containing a T2A-linked mCherry marker. (B) Analysis of luciferase activity of X1/5 cell pools (ON) transfected with either tetTALE or tetTALE-SD. P_tet-mediated luciferase activation of the TALE-negative parental cell lines with bound tet activator only was set to 100. Shown are mean values of three independent experiments with standard deviation. (C) Clones isolated from tetTALE-SD-transfected X1/5 cells were analyzed for tetTALE-SD linked GFP expression along with untransfected X1/5 cells (w/o) and the originating pool. (D) Single clones isolated from tetTALE-SD-transfected X1/5 cells grown under ON conditions were analyzed for luciferase activity 7 days after the switch of dox conditions from ON (-dox) to OFF (+dox). P_tet-mediated luciferase activation of the TALE-negative parental cell lines with bound tet activator only was set to 100. (E) Immunoblot analysis of single clones isolated from tetTALE-SD-transfected X1/5 cells grown under ON conditions to monitor tetTALE-SD expression levels. β-actin levels served as loading control.