MFP dose response in vitro
COS-1 cells were transfected with a 1:1
mixture of pGS1158 (CMV-GeneSwitch v3.1)/pAP1205 (6xGAL4/TATA-SEAP) or pGS1382
(4xGAL4/tk-GeneSwitch v3.1)/pAP1205, and treated with MFP at concentrations from 10 -13 to 10 -7 M. Induction of SEAP activity by pGS1158 occurred when MFP was administered at concentrations between 10 -12 and 10 -10 M, with half maximal induction at 10 -11 M. By contrast, induction of SEAP by pGS1382 occurred at MFP concentrations between 10 -11 and 10 -9 M, with half-maximal induction at 10 -10 M. Thus, the auto-inducible GeneSwitch® system required a higher concentration of ligand for transgene induction.
Figure legend.
MFP dose response curve was generated by transfecting COS-1 cells with a 1:1 mixture of
pGS1158 (CMV-GeneSwitch v3.1)/pAP1205 (6xGAL4/TATA-SEAP) (closed squares) or pGS1382
(4xGAL4/tk-GeneSwitch v3.1)/pAP1205 (closed circles). Twenty-four hours after transfection
the media was removed and replaced with media containing MFP at concentrations between 10 -13
and 10 -7 M. Cell culture supernatant samples were taken 24 hours later and
assayed for SEAP activity. The maximal induction of SEAP with pGS1158 (9-fold) and pGS1382
(188-fold) was at 10 -9 M MFP and is indicated above both curves. Data are
mean values ± SD of six independent transfections.
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