Fig. 4.

Greater bleb protrusion formation disrupts PCP and directed migration. (A) Live embryo and DIC control images (reproduced from Fig. 2A and Fig. 1A, respectively) of lateral mesodermal cells at the ypc/tb stage. Control and wild-type embryo injected with ezrb MO. White boxes indicate approximate position of microscopic analysis. DIC images are oriented as shown in Fig. 2A with dorsal to the right and anterior to the top. Select cells are outlined to show elongation and alignment relative to the dorsal-ventral body axis. D, dorsal. Scale bars: 5 µm. (B) Percentage of cells forming bleb protrusions during the 15 min time-lapse imaging (wild type at 80% epiboly+ca-ezrb, n=560 cells, 4 embryos; wild type at ypc/tb+ezrb MO, n=400 cells, 4 embryos; wild type at ypc/tb+ezrb MO+ca-ezrb mRNA, n=140 cells, 5 embryos). (C) PCP quantitation of lateral mesendodermal and mesodermal cells (wild type at 80% epiboly, n=150 cells, 4 embryos; wild type at 80% epiboly+ca-ezrb, n=126 cells, 4 embryos; wild type at ypc/tb, n=265 cells, 8 embryos; wild type at ypc/tb+ezrb MO, n=137 cells, 4 embryos; wild type at ypc/tb+ezrb MO+ca-ezrb mRNA, n=93 cells, 4 embryos). (D) MLA data from C depicted as individual data points (in degrees). Long black line indicate 20°. (E) Cell migration tracks toward the dorsal body axis. Origins (black arrows) were standardized for comparison. (F) Directness values for individual cells (wild type at 80% epiboly, n=50 cells, 3 embryos; wild type+ca-ezrb at 80% epiboly, n=60 cells, 3 embryos; wild type at ypc/tb, n=44 cells, 3 embryos; wild type+ezrb MO at ypc/tb, n=77 cells, 3 embryos). Box plot (B) shows the interquartile dataset, the median and the data range. Scatter plots (D,F) show individual data points and average values. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; unpaired Student's t-test with Welch's correction, except D (Watson non-parametric two-sample U² test).