Fig. 3

In silico feedback control of gene expression in mESCs. Set-point (a–d) and time-varying (e, f) reference relay control experiments, using inducers (Doxy 1000 ng/mL and/or TMP 100 nM) as control inputs. In red the measured output (normalised DDmCherry fluorescence), in blue the control reference fluorescence, set at 50% (a–d) or 50–70% (e, f) of the average maximal induction values measured during the calibration phase (120 mins with continuous Doxy/TMP (a–c, e) or Doxy (d, f) administration). The time-lapse sampling time was set at 60 mins. The effectiveness of controlling transcription (a, d, f), protein stability (b) and both (c, e) was tested: cells received either media with Doxy (a, d, f) or Doxy/TMP (c, e) or TMP (b) when the measured fluorescence was below the control reference, and plain media (c–f) or TMP (a) or Doxy (b) supplemented media when the measured fluorescence was above the control reference. The control performance indexes for each control experiment are reported in Supplementary Table 3 and Note 2. In the experiments in d, f, cells were never pre-treated with the protein stabiliser TMP (i.e. they only received Doxy). Details about feedback control experiments are in Supplementary Note 2