Figure 1—figure supplement 1.

Figure 1—figure supplement 1. (A) Schematic of (i) bilayered larval zebrafish skin and (ii) laceration technique. (B) Lacerated tailfin over time from a larva 3 days post fertilization (dpf) expressing LifeAct-EGFP in basal cells (TgBAC(∆Np63:Gal4); Tg(UAS:LifeAct-EGFP); Tg(hsp70:myl9-mApple)). mpw: minutes post wounding. (B–F) are all maximum-intensity Z-projections of spinning-disk confocal images. (C) Individual cell from 3 dpf larva expressing LifeAct-EGFP mosaically in basal cells (TgBAC(∆Np63:Gal4) larva injected with UAS:LifeAct-EGFP plasmid at the 1-cell stage). Wound was to the right approximately 1–2 min earlier. (D) Cells in a lacerated tailfin over time from 3 dpf larva expressing LifeAct-EGFP in basal cells (TgBAC(∆Np63:Gal4); Tg(UAS:LifeAct-EGFP); Tg(hsp70:myl9-mApple)), approximately 1–2 min post wounding. Arrowheads: examples of individual actin-rich protrusions are followed over time. (E) Kymograph indicating the speed of basal cells at a given distance from the wound over time (N = 8 larvae). Line graphs show net displacement over space (right) and time (bottom) for each individual larva. See Methods and Figure 1—figure supplement 1 for details of motion tracking analysis. (F) Lacerated tailfin from larva expressing GCaMP6f in basal cells (TgBAC(∆Np63:Gal4) larvae injected with UAS:GCaMP6f-P2A-nls-dTomato plasmid at the 1 cell stage). mpw: minutes post wounding. Due to the large dynamic range in GCaMP intensity, these images were gamma-corrected with a gamma of 0.5 for display purposes. (G) Kymograph of GCaMP6f intensity, normalized by the coexpressed nuclearly localized dTomato intensity, and relative to the normalized intensity pre-wounding (F₀) (N = 7 larvae). (H) Line graph of normalized profiles of the average speed and GCaMP intensity over time, averaged over 300 µm of tissue closest to the wound. To emphasize comparison of the temporal relationship, profiles are rescaled to lie between 0 and 1 (in arbitrary units).