
Fig. 6. (A) Xanx-4 MO specificity in vitro; western blot analysis showing that Xanx-4 MO interferes with in vitro translation of Myc-tagged Xanx-4 mRNA (Myc-Xanx-4) and not of Myc-tagged Xsox17ß mRNA (Myc-Xsox17ß). Lane 1: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA. Lane 2: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA and 1 µg Xanx-4 MO. Lane 3: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA. Lane 4: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA, and 5 µg Xanx-4 MO. Lane 5: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA. Lane 6: 10 ng Myc-Xanx-4 and Myc-Xsox17ß mRNA and 10 µg Xanx-4 MO. Lane 7: no input mRNA or MO. (B) Xanx-4 MO specificity in vivo; western blot analysis using anti-Myc antibody showing that the Xanx-4 MO interferes with in vivo translation in Xenopus embryos of Myc-tagged Xanx-4 mRNA (Myc-Xanx-4), lanes 2-4, and not of Myc-tagged Xsox17ß mRNA (Myc-Xsox17ß), lane 8. One-cell stage Xenopus embryos were injected with Myc-Xanx-4 mRNA, Myc-Xanx-4 mRNA and Xanx-4 MO, Myc-Xsox17ß mRNA, Myc-Xsox17ß mRNA and Xsox17ß MO, and cultured to stage 9, 13 and 21. Lane 1: 0.5 ng Myc-Xanx-4 mRNA. Lane 2: 0.5 ng Myc-Xanx-4 mRNA and 5 ng Xanx-4 MO. Lane 3: 0.5 ng Myc-Xanx-4 mRNA and 10 ng Xanx-4 MO. Lane 4: 0.5 ng Myc-Xanx-4 mRNA and 20 ng Xanx-4 MO. Lane 5: uninjected control. Lane 6: 0.5 ng Myc-Xsox17ß mRNA. Lane 7: 0.5 ng Myc-Xsox17ß mRNA and 10 ng Xsox17ß MO. Lane 8: 0.5 ng Myc-Xsox17ß mRNA and 10 ng Xanx-4 MO.