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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.