Fig. 3

Inhibition of Hsp90 by 17-DMAG and of the mTor pathway by rapamycin interfere with PGC arrival at their target. (A) Images of 20 hpf embryos treated with DMSO as control, or with the Hsp90 inhibitor 17-DMAG. Arrowheads, ectopic PGCs. Asterisk, migration target. The PGCs express farnesylated EGFP on their membrane. (B) Scatter plot of the mean percentage of ectopic PGCs per embryo in the control and in embryos treated with the inhibitor. Data derived from three independent experiments; error bars represent SEM. Statistical analysis performed using the Mann-Whitney-test, p < 0.0001. N is the number of embryos analyzed. (C) and (D) representative 70 min tracks of migrating PGCs in 6–8 hpf embryos treated with DMSO or with 17-DMAG in C, or with 17-DMAG and rapamycin together in D. (E) and (F) Bar graphs showing quantification of dynamic migration parameters for PGCs in embryos treated with 17-DMAG in E, or with 17-DMAG and rapamycin in F. Statistical analysis performed with stratified Kruskal-Wallis-test; for E displacement: p = 0.0002925; speed: p = 2.882e⁻⁰⁷; straightness: p = 0.1252 and for F displacement: p = 4.358e⁻⁰⁹; speed: p ≤ 2.2e⁻¹⁶; straightness: p = 0.007162. n.s. = not significant. n is the number of cells analyzed. (G) and (H) Proportion of the time PGCs exhibit polar morphology and active migration in 17-DMAG in G and in the combined treatment with 17-DMAG and rapamycin in H compared with the DMSO control. Error bars represent SEM. Data derived from three independent experiments. P-value for G: p = 0.0071 and for H: p < 0.0001, with Mann-Whitney-test. n is the number of cells analyzed.