Journal: bioRxiv

Article Title: Wunen(s) help navigate Primordial Germ Cells by attenuating Hedgehog signaling

doi: 10.64898/2026.05.01.722161

Figure Lengend Snippet: (A-B) Either UAS-lexA RNAi or UAS-wun RNAi males were mated with nos-gal4 . virgin females carrying both the transgenes were mated with endogenous Hh-GFP males. Compared to control lexA M- (A), wun M- embryos show a higher number of Hh-GFP puncta (Yellow) in the mesoderm. (C) Individual Hh-GFP puncta were counted in 15 ROIs dorsal to the gut, per genotype (gut outline is marked in all the panels). The increase in Hh-GFP is quantified using an unpaired T-test, ****p<0.0001. (A1-B1). Magnified insets shown in (A1’-B1’), with arrowheads marking Hh-GFP puncta. (D-F) Compared to control, lexA M- (D) wun M- embryos show a higher number of Hh-GFP puncta in their PGCs (E and F), marked with anti-Vasa antibody (red) staining. Magnified insets (D2’-F2’), with arrowheads marking Hh-GFP puncta. (G) The total number of puncta was counted in 15 PGCS per genotype across three replicates, and the percentages of PGCs with 1-2, 3-5, 6-8 and 9+ Hh-GFP puncta are plotted. Significance was calculated using Two-way ANOVA and Tukey’s multiple comparison test (****p<0.0001, ***p<0.001, **p<0.01).

Article Snippet: The following primary antibodies in specified dilutions were used: Rabbit anti-Vasa (1:1000, Gift from Trudi Schüpbach), Mouse anti-En (1:50, DSHB), Mouse anti-Wg (1:20, DSHB), Chicken anti-GFP (1:500, Invitrogen), Mouse anti-Smo (1:50, DSHB), Mouse (1:20), and Chicken anti-βgal (1:1000, Abcam).

Techniques: Control, Staining, Comparison

Journal: bioRxiv

Article Title: Wunen(s) help navigate Primordial Germ Cells by attenuating Hedgehog signaling

doi: 10.64898/2026.05.01.722161

Figure Lengend Snippet: All embryos shown are at Stage 10 (A-D) PGCs in control lexA M- embryo (A) exit the gut through its side and enter the mesoderm. In contrast, PGCs in wun M- embryos either clump precociously (B) or are delayed while migrating through the gut (C). PGCs of wun Z- (D) show a similar phenotype, although, as expected, the penetrance is lower. (E) PGC migration defects on depleting wun are quantified (n=15, 3 replicates) and significance determined using Ordinary one-way ANOVA and Tukey’s multiple comparison test (n=15, 3 replicates), ****p<0.0001, ***p<0.001. (F-G) wun2 M- leads to a highly penetrant precocious clumping phenotype in stage 10 (E) . wun2 Z- embryos also show similar phenotype (F) although not as penetrant. Arrowheads mark mis-migrated PGCs. Additionally, dying PGCs, marked by a star, are observed (H) . PGC migration defects on depleting wun2 are quantified (n=10, 2 replicates) and significance determined using Ordinary one-way ANOVA and Tukey’s multiple comparison test (n=10, 2 replicates) **p<0.01. Hatches represent the percentage of embryos that have PGCs that have not individualized by stage10. Embryos were stained with Hoechst to mark DNA (A1-D1 and E1-F1) and anti-Vasa (A2-D2 and E1-F1). (A3-D3 and E3-F3) display merged images.

Article Snippet: The following primary antibodies in specified dilutions were used: Rabbit anti-Vasa (1:1000, Gift from Trudi Schüpbach), Mouse anti-En (1:50, DSHB), Mouse anti-Wg (1:20, DSHB), Chicken anti-GFP (1:500, Invitrogen), Mouse anti-Smo (1:50, DSHB), Mouse (1:20), and Chicken anti-βgal (1:1000, Abcam).

Techniques: Control, Migration, Comparison, Staining