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Fig. 4. E-cadherin is dynamically regulated during early stages of PGC development. (A) The expression level of E-cadherin protein is altered during early development in wild-type PGCs (top row, blue, the contours of the relevant cells are delineated with white dots), but remains constant in PGCs knocked down for Dead end (bottom row). (B) A graphic representation of the quantitative analysis examining the relative E-cadherin level on PGCs. Regions in PGC and somatic cell membranes were selected for analysis and the average pixel intensity obtained from the PGC membrane (the inner half of the membrane, red) was divided by the average pixel intensity of the somatic cell membrane (the inner half of the membrane, yellow). (C) Wild-type PGCs at 5.3 hpf show significantly reduced E-cadherin levels relative to earlier stages (comparing 5.3 hpf with 3 hpf). dnd-MO treated PGCs showed no such change in E-cadherin levels on the membrane. (D) PGCs in which full-length E-cadherin is forced expressed show a strong reduction in cell motility (e.g. the cell marked with the green arrow). The PGCs exhibit extensive non-polarized protrusive activity (high magnification snapshots). (E) PGC migration speed is severely reduced compared with wild-type PGCs. (F) Embryos forced expressing E-cadherin (122 embryos examined) in PGCs show 30% ectopic germ cells in comparison to 5% of control embryos (120 embryos examined). (G) In 22 hpf embryos in which E-cadherin is forced expressed in the PGCs, ectopic PGCs can be observed (black arrowheads, cells labelled using a GFP probe). In (C,E) n is the number of cells analysed, the error bars represent the standard error of the mean (s.e.m.), asterisk signifies P<0.001.
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