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. 2011 Aug 18;6(8):e23734. doi: 10.1371/journal.pone.0023734

Figure 3. Easy generation of highly responsive cell lines with the all-in-one doxycline-controlable lentiviral vector CEST.

Figure 3

A). Schematic representation of the CEST. eGFP transgene is expressed from a Tetracycline –responsive CMV-TetO promoter and the TetR repressor is expressed from the SFFV promoter. B) Doxycycline responsiveness of 293 T containing different amounts of CEST vector copy per cell (0 (Mock), 0.4, 4 and 20 v.g.c. as indicated on the left hand side). The different cell lines were incubated in the absence of doxycycline (-Dox), and with 0.001 µg/ml, 0.01 µg /ml and 0.1 µg /ml as indicated on the top. The leaking in the absence of doxycycline decrease as the vector copy number increase (left graphs from top to bottom) C) Fold induction (left panels) and leaking (right panels) in 293 T (top panels) and human mesenchymal stem cells (hMSC) (bottom panels) transduced with increasing MOIs of the CEST vector. The average CEST v.g.c. of the different cell lines analyzed are indicated at the bottom of the graphs. The best regulation in terms of higher inducibility and lower leaking is achieved in the cells that contain the highest number of CEST vector integrated.