Effect of HDAC inhibition on the steady‐state and dynamic transcriptional response and associated noise. Related to Fig 7 Effect of HDAC inhibition on.

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Volume 104, Issue 4, Pages (February 2013)

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Effect of HDAC inhibition on the steady‐state and dynamic transcriptional response and associated noise. Related to Fig 7 Effect of HDAC inhibition on the steady‐state and dynamic transcriptional response and associated noise. Related to Fig 7 HDAC inhibitors have a dose‐dependent effect on transcriptional activity. MCF7‐GREB1‐PP7 cells were cultured with 100 pM E2 for 3 days, and then, butyrate (NaBu) or TSA was added for 4 h prior to fixation. Mean transcription sites intensities of > 5,000 cells per condition were calculated and fitted to a Hill function. Error bars denote standard deviation from two replicates. An intermediate butyrate concentration (*) was chosen for E2 titration experiments in panel (B).Inhibitors of deacetylation alter the intensity of transcription sites. MCF7‐GREB1‐PP7 cells were cultured at various concentrations of E2 for 3 days, after which 2.5 mM butyrate (NaBu) was added for 4 h. Transcription sites were detected automatically and quantified using > 2,500 cells per condition. The mean and standard deviation from two biological replicates are plotted and fitted to a Hill function.Intensity histograms of HDAC inhibitor titration reveal qualitative differences in noise behavior as compared to the E2 dose‐response. Kernel density estimates were calculated from spot intensities from high‐content imaging datasets (same data as in panel A and B). Comparison with stochastic simulations (panel D) indicates a decrease in burst size at low NaBu concentrations, as compared to the E2 dose‐response. Low NaBu doses primarily affect intensity but not the frequency, of the right peak in the histogram (see arrow).Simulated intensity histograms distinguish burst size from burst frequency modulation. Stochastic simulations were performed (tON = 1 min; b = 8 RNAs/burst; model topology: 1–1–5) with OFF‐times ranging from 10 (yellow) to 100 min (blue) and global intensity histograms were calculated. Both the proportion and intensity of active transcription sites changes with OFF‐time (top). Burst sizes were changed by increasing the initiation rate from 1 (blue) to 40 RNAs/min (yellow) during stochastic simulations (tON = 1 min; tOFF = 30 min; model topology: 1–1–5) (bottom). Only the intensity of active transcription sites (x‐position of the right peak) changes with changing burst sizes. Christoph Fritzsch et al. Mol Syst Biol 2018;14:e7678 © as stated in the article, figure or figure legend