Extended Data Fig. 7: Investigating the role of Mediator in spatiotemporal confinement of RNAPII. | Nature Cell Biology

Extended Data Fig. 7: Investigating the role of Mediator in spatiotemporal confinement of RNAPII.

From: Disordered C-terminal domain drives spatiotemporal confinement of RNAPII to enhance search for chromatin targets

Extended Data Fig. 7

a, Bound Mediator map (left). Free RNAPII-CTD26 map showing confined trajectories (right). Scale bar: 0.5 μm. (Error bar: centroid of nucleolus ± s.d.; n: number of nuclei). b, Free RNAPII-CTD26 map in MED14Rgr1 AID. Scale bar: 0.5 μm (Error bar: centroid of nucleolus ± s.d.; n: number of nuclei). c-d, c, Dfree in nucleoplasm (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, ns P = 0.510991) and d, Nucleoplasm/Nucleolus of RNPAII-CTD26 in MED14Rgr1 AID (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, *** P = 0.000008). e-g, e, Mediator bound fraction (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, ** P = 0.006771), f, RNAPII bound fraction (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, ns P = 0.157495) and g, f180/0 for RNAPII-CTD9 in CTD26 background (n = 100 resamplings; mean value ± s.d.). h,i, h, Relative bound fraction (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, * P = 0.012696, ** P = 0.001329) and i, f180/0 for RNAPII-CTD26 and RNAPII-CTD9 in WT and ssn3Δ (n = 100 resamplings; mean value ± s.d.). Deleting Cdk8SSN3 from the Mediator kinase module enhances interaction between the Mediator and RNAPII97,98,99 without changing free RNAPII confinement. j,k, j, Relative bound fraction (n = 100 resamplings; mean value ± s.d.; two-tailed unpaired t-test, * P ≤ 0.05 and *** P ≤ 0.001. Comparison between before and after auxin treatment: P = 0.000014 for CTD26 and P = 0.0222222 for CTD9. Comparison between CTD26 and CTD26 after auxin treatment: P = 0.034745) and k, f180/0 (n = 100 resamplings; mean value ± s.d.) for RNAPII-CTD26 and RNAPII-CTD9 in MED14Rgr1 AID. l–n, l, Survival probability of H2B-corrected residence times (n = 10,000 resamplings; mean value ± s.d.), m, stably bound fractions (n = 10,000 resamplings; mean value ± s.d.; two-tailed unpaired t-test, ** P = 0.002245) and n, mean residence times (n = 10,000 resamplings; mean value ± s.d.; two-tailed unpaired t-test, * P = 0.019928) for RNAPII-CTD9 in MED14Rgr1 AID. Source numerical data are available in source data.

Source data

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