A. Whole mount in-situ hybridisation (WISH) in early embryos shows altered expression of germline markers ddx4, dnd1, and nanos1 in igf2bp3Δ7 mutant embryos. B. qRT-PCR to detect early germplasm markers shows reduced levels of ddx4 and nos1 expression in mutant embryos at the 4-cell stage, whereas expression levels of dnd1 is not significantly different from control wild type embryos. C. Primordial germ cells (PGCs; red arrowheads) in igf2bp3Δ7 embryos are ectopically located at the 1k-cell stage to varying extents ranging from mild or moderate to severe. D. Primordial germ cells are severely reduced or not detected in igf2bp3Δ7 mutants by gastrula stages. E,F. WISH (E) and quantitation (F) of ddx4-positive cells (red arrowheads) shows reduced and ectopic primordial germ cells in 24 hpf maternal igf2bp3Δ7 (Migf2bp3Δ7) and maternal-zygotic igf2bp3Δ7 mutants (MZigf2bp3Δ7) compared to WT siblings and paternal igf2bp3Δ7 (Pigf2bp3Δ7) mutants; p *<0.05, **<0.01, ***< 0.001. G. Loss of maternal igf2bp3 leads to some ectopic primordial germ cells across the trunk (red arrowheads) and occasionally in the hindbrain region. Bar graph shows the number of embryos with ectopic germ cells in WT, Pigf2bp3Δ7, Migf2bp3Δ7, and MZigf2bp3Δ7 mutants. Scale bar in A and C-E, 200 μm. N = 15 embryos each for WT, Pigf2bp3Δ7, and Migf2bp3Δ7 and 39 for MZigf2bp3Δ7 mutants.
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