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Fig. 6. Depletion of XlPouV proteins affects germ layer induction. (A) Two-cell stage embryos were injected in both blastomeres with 60 ng of control MO or 20 ng of each of the XlpouV MOs. In situs were performed on stage 10.25 embryos. Embryos are positioned with the vegetal hemisphere upwards and the dorsal lip towards the top apart from Bmp4 and Xom, which are positioned animal hemisphere upwards and insets with vegetal hemisphere upwards. (B) Real-time RT-PCR analysis of XlpouV protein-depleted embryos. Embryos were injected with control MO or XlpouV MO as in Fig. 6A and RNA extracted at stage 10.25. The relative change in gene expression was calculated by dividing the XlpouV MO values by the control MO values. Experiments were carried out in triplicate. (C) Embryos were injected with control MO or XlpouV MO as in Fig. 6A. Sox2 and Ngnr1 expression was analysed at stage 13 and Hex and MLC1-3 expression at stage 35 embryos. (D) Two-cell stage embryos were injected in both blastomeres with 250 pg of each RNA and in situs performed on stage 10.25 embryos. All embryos are positioned with the vegetal hemisphere facing upwards and the dorsal lip towards the top, apart from Bmp4 embryos that are positioned animal hemisphere upwards. |
