Fig. S5
Demonstration of the efficacy and specificity of the E-cadherin MO. (A) Embryos injected with 300 pg of E-cadherin MO exhibit decreased E-cadherin levels by 24 hpf, whereas wildtype embryos and those injected with 300 pg of a five-base mismatch E-cadherin MO do not. A representative immunoblot is shown, and quantitative data are the average E-cadherin/β-actin ratios of at least three independent samples ± s.e.m., normalized with respect to the wildtype condition. (B–C) Distribution of vasa-positive PGCs in embryos injected with either water or the mismatch E-cadherin MO (300 pg/embryo) at the one-cell stage and then treated with 50 μM cyclopamine until 24 hpf. Scale bar: 200 μm. (D) Quantification of PGC mislocalization in embryos injected with either water or the mismatch E-cadherin MO (300 pg/embryo) and treated with 50 μM cyclopamine or an ethanol vehicle control. At least 600 PGCs were scored for each experimental condition. |
Reprinted from Developmental Biology, 328(2), Mich, J.K., Blaser, H., Thomas, N.A., Firestone, A.J., Yelon, D., Raz, E., and Chen, J.K., Germ cell migration in zebrafish is cyclopamine-sensitive but Smoothened-independent, 342-354, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.