ZFIN Image: Georgantzoglou et al., 2022, Fig. 5

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Figure Caption

Fig. 5

Differential contributions of Arp2/3 and Myosin-II in gradient responses. (A) Top row: Two-photon time-lapse image projections showing neutrophils in Tg(mpx:Lifeact-Ruby) zebrafish larvae treated with DMSO, CK666, and Blebbistatin, where the cell front has been segmented automatically (red lines). Bottom row: Example neutrophil treated with Blebbistatin, with new leading edge forming at the cell lateral/rear towards the LW (red arrow). White arrow indicates the cell’s direction of migration. Triangle indicates the direction of LW (not in the field of view). Scale bar = 10 μm. (B) Average surface area of the cell front part with DMSO, CK666, and Blebbistatin treatments. One-way ANOVA with Dunnett’s multiple comparisons test, * P = 0.0138 (DMSO-CK666 pre-LW), P = 0.0437 (CK666-Blebbistatin pre-LW), P = 0.0379 (DMSO-CK666 post-LW). (C) Percentage of cells that generate a new leading edge from the rear or the side of their body, for the treatments of DMSO, CK666, and Blebbistatin, respectively. Chi-square test, ****, P < 0.0001. (D) Average time that cells spend in search (front Lifeact enrichment) and running (rear Lifeact enrichment) phase, for the treatments of DMSO, CK666 and Blebbistatin, respectively, for pre- (left) and post-LW (right). Kruskal-Wallis test with Dunn’s multiple comparison test, ***, P = 0.0005 for time in search mode, *, P = 0.0124 for time in running mode. (E) Percentage of cells with persistent steps (θ < 60° for δ < 45°). n = 9 larvae for DMSO, n = 5 larvae for CK666, and n = 4 larvae for Blebbistatin. Wilcoxon matched-pairs signed rank test, *, P = 0.0195 for DMSO, P = 0.0216 for Blebbistatin. (F) Mean instantaneous speed for movement towards (TW) or away (AW) from the wound, for pre- and post-LW. n = 1,334–2,292 cell steps per condition for DMSO (9 larvae), n = 1,056–1,616 cell steps per condition for CK666 (5 larvae), and n = 818–1,826 cell steps per condition for Blebbistatin (4 larvae). Kruskal-Wallis test with Dunn’s multiple comparison test, ****, P < 0.0001 for both DMSO and CK666. (G) Persistence of cells for 3 min (left) and 6 min (right) after neutrophil movement for DMSO, CK666, and Blebbistatin. One-way ANOVA with Dunnett’s multiple comparisons test, **, P = 0.0004 (3 min, DMSO-Blebbistatin), *, P = 0.0154 (6 min, DMSO-CK666), ***, P = 0.0002 (6 min, DMSO-Blebbistatin). (B–D and G) Data from n = 17 cells from 8 larvae for DMSO, n = 11 cells from 5 larvae for CK666, and n = 13 cells from 4 larvae for Blebbistatin. Mean and SEM are shown.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ J. Cell Biol.