Fig. S4

Fig. S4

The role of vesicle trafficking in blebbing. Related to Figure 4

(A) Labeling of the Golgi apparatus in a migrating PGC using human beta 1, 4- galactosyltransferase target sequence fused to dsRed (dsRed-Gtransferase). The plasma membrane is labeled with EGFP-F or with mCherry-F (presented in green). Arrow shows the direction of migration and the position of the nucleus is outlined with the dashed-lines. Localization of different vesicle markers including VAMP8, Rab11a, Rab5c and Exo70 and their position relative to the forming bleb (asterisk). (B) 24 hpf embryos globally expressing control RNA or dominant negative (DN) forms of Exo70, Rab11a and Rab5c (upper two rows) and 24 hpf embryos expressing the same dominant negative forms preferentially in the PGCs by employing the nanos3 3’UTR (lower two rows). In all cases, the PGCs reach their clustering position. (C) PGCs expressing the dominant-negative of Exo70, Rab11a and Rab5c show a frequency of bleb formation similar to that of control cells. Scale bar=5 µm. Statistical analysis was conducted using the Kruskal-Wallis test, with P value= 0.1, ns. The graph presents the combined data from 3 independent experiments. The graphs show median and interquartile range. Ctrl stands for Control RNA.