Supplementary Figure 14: Dose dependence of the small-molecule–responsive transcription-control systems used for the assembly of logic gates.
From: A general design strategy for protein-responsive riboswitches in mammalian cells
(a) HEK-293 cells cotransfected with the vanillic acid-dependent transrepressor (VanA4; pCK188) and the VanA4-specific PvanON8-driven SEAP expression vector with embedded Env140/nutR/H6ac (PVanON8-SEAP-pA; pSA783) and either N-peptide-mCherry (pSA776; circles) or mCherry (pFS29; squares) were programmed with different concentrations of vanillic acid and profiled for SEAP expression after 40h. (b) HEK-293 cells were cotransfected with the reverse tetracycline-dependent transactivator (rtTA; pTet-ON) and the rtTA-specific Ptight-driven N-peptide-mCherry expression vector (Ptight-N-peptide-mCherry-pA; pSA781) and programmed with different concentrations of the antibiotic doxycyline. Red fluorescence was quantified by flow cytometry 40 hours after transfection. (c) HEK-293 cells cotransfected with the erythromycin-dependent transactivator (ET1; pWW35) and the ET1-specific PETR2-driven driven N-peptide-mCherry expression vector (PETR2-N-peptide-mCherry-pA; pSA763) and programmed with different concentrations of the antibiotic erythromycin. Red fluorescence was quantified using flow cytometry 40 hours after transfection. Error bars are mean±SEM (n=3).
