Fig. 4

Tdrd6a Is Required for Gp Integrity

(A) Confocal images of Buc-eGFP-positive embryos from tdrd6a^+/− mothers, at 1-, 2-, and 4-cell stages, focusing on Gp as schematically indicated. In zoom of the Gp, IHC for Tdrd6a and double smFISH was performed and displayed as indicated. Arrowheads indicate mRNA and Tdrd6a that is peripherally localized on the Gp granule; arrows indicate Buc-eGFP bridges (zooms 1 and 2 on the right). Asterisks mark mature Gp, containing fused Buc-eGFP and mRNA networks inside the structure. Line graphs display intensity for Buc (green) versus Tdrd6a (red) signals over line a-b and dazl (red) versus vasa (cyan) intensity over line c-d, with vertical lines indicating fluorescence peaks per smFISH signal (highlighted by colored circle on top).

(B) Confocal images of Buc-eGFP-positive Gp of a 4-cell stage embryo from a tdrd6a^−/− mother (Mmut). SmFISH was performed and displayed as indicated. Arrows indicate areas where Buc-eGFP has fused, but mRNA remains peripherally localized.

(C) Boxplot representing volumes of the Buc-eGFP signal at the cleavage planes of wt and Mmut 4-cell stage embryos. The largest Gp fragment of each embryo was measured (also see Figure S5B) after 3D reconstruction in Imaris. (^∗ indicates p value < 0.001, Wilcoxon test).

(D) Quantification of Gp phenotypes of 4-cell stage embryos using an ISH against vasa. Scale bars, 10 μm (overview A), 2 μm (zoom) (A), and 2 μm (B).

See also Figure S5.