Fig. 3. A circadian rhythm in fibroblast wound-healing response.

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Fig. 3. A circadian rhythm in fibroblast wound-healing response. A circadian rhythm in fibroblast wound-healing response. (A) (i) Fibroblast monolayers derived from adult PER2::LUC mouse skin were entrained and wounded after 20 to 64 hours in free run. (ii) Images of wound-healing assays; time at wounding is indicated, and residual wound is indicated by the highlighted area (pink) (scale bar, 500 μm). (iii) Quantification of the residual wound after 16 hours of wound healing (line) (n = 4; mean ± SEM) with three parallel PER2::LUC measurements (heat maps). RAIN P value is indicated. (B) (i) Fibroblasts labeled with CellTracker Red healing after wounding at the indicated times after synchronization (t). Scale bars, 100 μm. Wound healing (ii) and leading cell velocity (iii) are quantified (n = 4 or 5; mean ± SEM). P values from Tukey’s multiple comparisons test after 60 hours of healing (tH = 60) (ii) or tH = 0 (iii) are indicated. (C) (i) Fibroblasts labeled with SiR-actin (red) and CellTracker Green (cyan) treated as in (B). Scale bars, 50 μm. A single cell for each condition has been highlighted in white with time healing indicated. The center of mass for each label (ii) was determined over 9 hours of healing, and the mean ± SEM degree of polarization (Δx) is indicated (iii). The P value from a two-way analysis of variance (ANOVA) is indicated. Nathaniel P. Hoyle et al., Sci Transl Med 2017;9:eaal2774 Published by AAAS