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Fig. 4. XTgfbi was required for dorsal tissue specification and differentiation. Whole-mount in situ hybridization of embryos with mis-expressed XTgfbi. The injected reagents, probes, and stage of embryos are indicated in the lower right, left, and upper-right corners, respectively. The injected sides are marked by β-gal. (A) XTgfbi depletion inhibited MHC expression in whole mount and sectioned embryos at stage 21. The dashed lines indicate the bent midline. In the longitudinal sections, the injected sides are indicated by the arrows. (B) XTgfbi-depletion repressed MyoD expression in stage 20 and 11 embryos. (C, D) hTgfbi or XTgfbi-MT injection rescued the loss of MHC caused by MO1 or MO2. Summaries of MHC expression based on arbitrary scores analyzed with unpaired two-tailed Student T test (P=0.009 hTgfbi, P=0.032 XTgfbi-MT). Error bars represent s.e.m. C: Representative MHC stained embryo injected with 2 ng hTgfbi only (n=21), 2 ng hTgfbi with 34 ng MO1 (n=29) and 34 ng MO1 only (n=33), respectively. D: Representative MHC stained embryos injected with 2 ng XTgfbi-MT only (n=22), 2 ng XTgfbi-MT with 34 ng MO2 (n=19) and 34 ng MO2 only (n=14), respectively. (E) XTgfbi depletion inhibited NBT expression at stage 15 and 21. The red bars indicate the neural plate expansion. (F) XTgfbi-depletion expanded neural plate marker Sox2 at stage 15. (G) XTgfbi-depletion repressed the expression of neural crest markers Twist at stage 21. (H) Cartilage staining of representative uninjected and MO1-injected embryos at stage 42. Embryos with XTgfbi-depletion developed smaller cranial cartilage.
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