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FIGURE 6. Jmjd6 is required for Xenopus embryonic development. A, Co-IP detection of the
interaction between Jmjd6 and Tcf7l1 in Xenopus embryos. Each 300 pg of mRNA for tagged Jmjd6
or/and Tcf7l1 was injected into 2-cell stage embryos. At gastrula stage, embryos were collected and
subjected to Co-IP assays. B, design of an antisense morpholino oligonucleotide, Jmjd6MO, against both
the transcripts of jmjd6a and jmjd6b. Translational start site of each transcript is underlined. C,
immunoblotting detection of the knockdown efficiency of the Jmjd6MO, as compared with the standard
control morpholino (ctrlMO). One nanogram of mRNA for HA-tagged Jmjd6a that contains the
Jmjd6MO binding site was injected alone or co-injected together with 20 ng of either ctrlMO or
Jmjd6MO into Xenopus embryos. Embryos were collected at gastrula stage and subjected to
immunoblotting. D, typical Xenopus embryos after injection of ctrlMO or different doses of Jmjd6MO,
showing severe developmental defect in response to Jmjd6 knockdown. The defect became stronger in
response to a higher dose of injected Jmjd6MO. Embryos are shown in lateral view, with the anterior
being placed to the right of each panel. E, numbers and percentages of total and defect embryos in the
experiments in (D). F, different changes in the expression of genes that are involved in anterio-posterior
body axis patterning in gastrula (for cer1, dkk1, gsc and chrd) and neurula (for xag2) embryos after
injection of 20 ng of ctrlMO or Jmjd6MO. v: vegetal view, with the dorsal being orientated to the top of
each panel; a: anterior view, with the dorsal being placed to the top. G, numbers and percentages of
embryos showing different changes in gene expression after Jmjd6 knockdown. |
