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Fig. 6. Sox9 depletion leads to a loss of neural crest progenitors and an expansion of neural tissues. (A) Embryos were injected in one blastomere at the two-cell stage with 10 ng of Sox9-AS or Co-AS and analyzed by whole-mount in situ hybridization at stage 17-19 (Snail, Pax3, Nrp1 and Sox2) or stage 23 (Twist). Dorsal view, anterior is at the top. RNA encoding the lineage tracer β-galactosidase was co-injected to identify the injected side (red staining), the right side in all panels. Upon injection of Sox9-AS, expression of the neural crest markers Twist, Snail and Pax3 are greatly reduced, while expression of the pan-neural marker Nrp1 and Sox2 is expanded. Expression of the same markers in Co-AS-injected embryos is presented for comparison. (B) Tissue sections of representative Sox9-AS-injected embryos stained for Slug or Sox2 expression. Injected side is on the right. n, notochord; s, somite. |
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Fig. 7. The phenotype of Sox9-depleted embryos can be rescued by restoring Sox9 expression. (A) Rescue experiments were performed by injection of an animal dorsal blastomere at the eight-cell stage. (B) Representative case of Slug whole-mount in situ hybridization of stage 17 embryos injected with 100 pg of Sox9 plasmid (Sox9) or 5 ng of Sox9-AS (Sox9-AS), or a combination of both (Sox9-AS+Sox9) or a combination of Sox9-AS and 100 pg of a control GFP plasmid (Sox9-AS+GFP). RNA encoding the lineage tracer β-galactosidase was co-injected to identify the injected side (red staining), right side in all cases. Dorsal view, anterior is at the top. (C) Quantification of the in situ hybridization results. n, number of cases analyzed. |
