Fig. 7: Moesin is required to regulate hemocyte polarity, leading edge dynamics, and actin retrograde flow.

a Morphodynamic analysis of hemocyte polarity. b Quantification of polarity highlighting a reduction in the absence of Moesin. *P  =  0.0332, Mann–Whitney two-tailed test. (n = 13 Control and 14 Moesin^Mut hemocytes). c Dynamic analysis of leading edge velocities highlighting the edge velocity of all extensions (green) or the largest contiguous extension (blue). Extension vector magnitude has been multiplied by 10 for visualization purposes. d Quantification of speed of the largest extension or all extensions highlighting a reduction in the absence of Moesin. ****P  <  0.0001, Mann–Whitney two-tailed test. (n = 677 Control and 995 Moesin^Mut timepoints for 13 and 14 hemocytes, respectively). e Comparison of the direction of cell motion (magenta) with the direction of the largest extension (blue) in Control and Moesin^Mut hemocytes. f (left panel) Quantification of the correlation in motion (cos(theta)) between the largest extension (blue) and the direction of movement (magenta) in Control and Moesin^Mut hemocytes highlighting a decreased correlation in the absence of Moesin. (right panel) Rose plot comparing the direction of the largest extension normalized to the instantaneous hemocyte direction of motion. ***P  = 0.0008, Mann–Whitney two-tailed test. (n = 677 Control and 995 Moesin^Mut timepoints for 13 and 14 hemocytes, respectively). g PIV analysis of actin flow in Control and Moesin^Mut hemocytes. h Quantification of actin flow speed in Control and Moesin^Mut hemocytes highlighting a reduction in the absence of Moesin. *P  =  0.0128, Mann–Whitney two-tailed test. (n = 18 Control and 14 Moesin^Mut hemocytes). (n = 18 Control and 14 Moesin^Mut hemocytes). i PIV analysis of actin flow when expressing two copies of Moesin^WT or Moesin^TD. j Quantification of actin flow speed when expressing two copies of Moesin transgenes revealing a reduction in retrograde flow in Moesin^TD cells, which is rescued when combining with a K/N mutation. *P  =  0.0286, Kruskal–Wallis test and Dunn’s multiple comparison test. (n = 18 Control, 11 Moesin^WT, 8 Moesin^KN, 9 Moesin^TD, 7 Moesin^TDKN, 8 Moesin^TA and 12 Moesin^TAKN hemocytes). All boxplots show medians, 25th and 75th percentiles as box limits, minimum and maximum values as whiskers; each datapoint is displayed as a marker. Scale bars, 10 µm.