Figure 4

Disruption of rab10 balance leads to PGC mislocalization. (A) qRT-PCR analysis of rab10 mRNA in 24hpf embryos injected with control siRNA (con siRNA) or gradient rab10 siRNA (50, 150, 300 pg). (B) Co-injection of rab10 siRNA decreased expression of RFP in embryos injected with rab10-rfp mRNA compared to embryos co-injected with control siRNA. All embryos were co-injected with pEGFP-N3 vector as a control. (C) Quantitative representation of the normalized signal intensity in the experiment presented in (B). (D–M) Representative images of embryos injected with 300 pg con siRNA (D), 50 pg rab10 siRNA (E), 150 pg rab10 siRNA (F), 300 pg rab10 siRNA (G), 200 pg rfp mRNA (H), 50 pg rab10 mRNA (I), 100 pg rab10 mRNA (J), 200 pg rab10 mRNA (K), rab10T23N mRNA (L), and rab10Q68L mRNA (M). Arrowheads indicate mislocalized PGCs. (N) The percentage of PGC phenotypes in embryos with (D–K). (O) The number of mislocalized PGCs in each embryo with the experiment presented in (D–K). The average number of mislocalized PGCs were showed in red line by mean ± sd. (A,C,N,O) The results were representative of more than three independent experiments in triplicate. β-actin was used as an internal control to normalize gene expression levels with 2^−∆∆Ct method. ** p < 0.01; *** p < 0.001, **** p < 0.0001. Scale bar, 100 µm.