Supplementary Materials

Supplementary Material for:

Circadian actin dynamics drive rhythmic fibroblast mobilization during wound healing

Nathaniel P. Hoyle,* Estere Seinkmane, Marrit Putker, Kevin A. Feeney, Toke P. Krogager, Johanna E. Chesham, Liam K. Bray, Justyn M. Thomas, Ken Dunn, John Blaikley, John S. O'Neill*

*Corresponding author. Email: nhoyle{at}mrc-lmb.cam.ac.uk (N.P.H.); oneillj{at}mrc-lmb.cam.ac.uk (J.S.O.)

Published 8 November 2017, Sci. Transl. Med. 9, eaal2774 (2017)
DOI: 10.1126/scitranslmed.aal2774

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Cell-intrinsic rhythms in the fibroblast proteome.
  • Fig. S2. Validation of cofilin 2 and RCC2 circadian abundance.
  • Fig. S3. Circadian clocks are robust against actin-modulating drugs.
  • Fig. S4. Cell-intrinsic rhythms in actin polymerization in NIH3T3 cells.
  • Fig. S5. Cell division in healing fibroblast monolayers.
  • Fig. S6. Cell area, velocity, and cortical actin distribution are not circadian in confluent cultures with circadian PER2::LUC activity.
  • Fig. S7. Circadian rhythms in actin dynamics and wound-healing efficacy are CRY-dependent in embryonic fibroblasts.
  • Fig. S8. Fibroblast monolayer healing in the presence of cytoD.
  • Fig. S9. Rho inhibition disrupts impedance rhythms and time-of-wounding effects without disrupting the circadian clock.
  • Fig. S10. Collagen deposition and fibroblast distribution after 14 days of healing.
  • Fig. S11. A model of clock control of wound healing.
  • Fig. S12. Circadian rhythmicity of the “Regulation of actin cytoskeleton” pathway.
  • Table S1. DAVID clustering of rhythmic proteins identified by RAIN (P < 0.01).
  • Legends for movies S1 to S4
  • References (4856)

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Other Supplementary Material for this manuscript includes the following:

  • Table S2. Individual subject-level data for n < 20 (provided as an Excel file).
  • Movie S1 (.mov format). Healing of fibroblast monolayers wounded 24 hours after synchronization.
  • Movie S2 (.mov format). Healing of fibroblast monolayers wounded 32 hours after synchronization.
  • Movie S3 (.mov format). Fibroblast invasion of wounds made in the active phase after 48 hours of healing.
  • Movie S4 (.mov format). Fibroblast invasion of wounds made in the inactive phase after 48 hours of healing.