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Fig. 4. XProfilin2 does not interfere with mesoderm induction. Embryos injected dorsally with Profilin2 (2 ng), XProfilin2 MO (70 ng), XProfilin2 MO + δN-XProfilin2 (70 ng + 100 pg) have no defects in expression of mesodermal marker genes Xbra, Gsc and XWnt8 as monitored by RT-PCR analysis; Ornithine decarboxylase (ODC) is used as a loading control, −RT without reverse transcriptase. (B) Embryos injected dorsally with XProfilin2 (2 ng), XProfilin2 MO (70 ng), XProfilin2 MO + δN-XProfilin2 (70 ng + 100 pg) show abnormal tissue localization due to gastrulation defects. Injected embryos show normal expression of Xbra at st 10.5 but Xbra is observed trapped around the blastopore that does not close at st 14. Sox-2 is expressed in the neural plate at st 14 in uninjected embryos, but in the injected embryos, Sox-2 expression surrounds the open blastopore. Gsc expression is observed in anterior mesendoderm away from the closed blastopore in control embryos at st 14, but in the injected embryos remains trapped near the open blastopore. Otx-2 is expressed anteriorly in both mesodermal and overlying neural tissues in control embryos at st 14, but in the injected embryos, the Otx-2 expression domain remains closer to the blastopore. Number of embryos scored is shown on each panel and yellow arrow indicates position of the blastopore. |
