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. 2018 Dec 27;47(4):e23. doi: 10.1093/nar/gky1286

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© The Author(s) 2018. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Demonstration of pMVP inducible systems. (A) Graphical depiction of the mechanisms for the Tet-On, FKBP/ecDHFR degron domain, and AID systems and (B) the pMVP vectors generated and utilized. (C) Immunoblot analysis of INS1 832/13 cell lysates transduced with crude lysates for adenoviruses expressing PDX1 using either the Tet-On, ecDHFR, or FKBP systems and harvested at the indicated time intervals after addition (induction) and removal (washout) of the respective Dox, TMP, or Shld1 inducers. (D) Immunoblot analysis of INS1 832/13 cell lysates transduced with crude lysates for adenoviruses expressing PDX1 using either the Tet-On or ecDHFR systems and harvested 24h after addition of a dose curve of Dox (0.5–100 ng/ul) or TMP (1 nM–5 uM), respectively. (E) Immunoblot analysis of cell lysates from an INS1 832/13-derived cell line made by Sleeping Beauty-mediated integration of Tir1-P2A-AID-PDX1. Lysates were harvested at the indicated time intervals after treatment with 500uM auxin. (C–E) Endogenous PDX1 and PDX1-fusion bands were distinguished based upon size or the use of an epitope tag and are labeled accordingly.