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Fig. 4 Gγ2C68S- and Gαt-expressing PGCs fail to respond to Cxcl12a. (A) Schematic illustration outlining the chemoattractant assay. cxcl12a(-/-) embryos at the one-cell stage were injected with gfp-nos mRNA (to label PGCs) alone (control) or together with mRNAs encoding either Gγ2C68S-nos1-3′UTR or Gαt-nos1-3′UTR. At the 64-cell stage, a single cell at the animal pole was co-injected with RNAs encoding Cxcl12a and mCherry (tracer). (B,C) Snapshots from time-lapse movies showing the positions of PGCs (green) and Cxcl12a-expressing cells (red) in control embryos (B, supplementary material Movie 6) or in embryos injected with Gγ2C68S-nos1-3′UTR RNA (C, supplementary material Movie 6) at 8-9 hpf. White arrows indicate protrusions. (D,E) Representative embryos at 24 hpf showing localization of control or Gαt-expressing PGCs (green) and the Cxcl12a-expressing cells (red). Inset indicates high-magnification image of the boxed area. Scale bars: 10 μm in B,C; 200 μm in D,E. (F) Quantitative analysis of PGC colocalization with Cxcl12a-expressing cells at 24 hpf. *P<0.01 versus control. Data are mean+s.e.m.