Fig. 4

Dnd Is Required for Maintenance of Germline RNA Expression and for Inhibition of Somatic RNA Expression

(A) Upregulation of the muscle-specific master regulator myoD and downregulation of the germline-specific marker vasa within Dnd-deficient PGCs, as determined by using RNAscope in situ hybridization. Confocal plane images (4.1 μm thick) were acquired at 24 hpf within myoD-expressing muscle precursors. Directional cell migration was inhibited by knockdown of Cxcl12a. Scale bars, 20 μm.

(B) Graph presenting the number of vasa- and myoD-positive RNA spots detected within PGCs in control embryos and embryos knocked down for Dnd (quantification of results as presented in A). p Values determined using Mann-Whitney U test with α = 0.05. Data are presented as median ± IQR.

(C) PGCs (red) captured in transgenic embryos expressing a muscle-specific transgene unc45b:tfp (blue) labeling muscle precursors in control and Dnd-deficient cells. Directional cell migration was inhibited by knockdown of Cxcl12a. Confocal plane images, 5 μm thick, captured at 24 hpf. Scale bars, 20 μm.

(D) Graphs presenting signal intensity ratios of the muscle-specific transgene (as in C). Signal intensities within the PGC (1) were compared to intensities in a neighboring area of the same shape (2). p Values determined using Student’s t-test with α = 0.05. Data are presented as median ± IQR.

n, number of cells; F′, farnesylated; KD, knockdown; hpf, hours post fertilization.