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Fig.4. Mesoderm-Specific Xpbx1 and Xhoxa1 Knockdown in Xenopus Embryos(A) Diagram of marginal zone V2.2 Xenopus blastomere injection at 16-cell stage, and fate of injected blastomere in stage 17 neurula.(B) Whole-mount detection of RFP in left V2.2 blastomere-injected embryo.(C) Merge of bright field and fluorescence pictures of the embryo in (B).(D) Cross-section showing selective RFP labeling in somites (s) and lateral plate mesoderm (lpm).(E–I) Whole-mount double in situ hybridization for Xraldh2 and Xkrox20 in control-MO (E), Xpbx1b-MO (F and G), and Xpbx1b-MO;Xhoxa1-MO (H and I) left V2.2 blastomere-injected embryos.(J) Nuclear-salmon-gal staining (red cells) of Xpbx1b-MO;Xhoxa1-MO V2.2-injected embryos indicate morphant cell localization in mesoderm. Xkrox20 expression shows r3* posteriorization and r5* loss on the injected side.(K and L) Correlation between Xraldh2 downregulation and hindbrain phenotype in Xpbx1b-MO and Xpbx1b;Xhoxa1-MO injected embryos (mosaic plots). Relative phenotype severity is color-coded. Phenotype frequencies (y axis) are compared to levels of Raldh2 (x axis). Synergistic action of Xpbx1b-MO;Xhoxa1-Mo results in rising the frequencies of Xraldh2 severe reduction or loss and hindbrain patterning defects. MO, morpholino; r, rhombomere; RFP, Red Fluorescent Protein.See also Figure S2. |
