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Figure 2. XRFC is required for neural crest induction and migration.
(A) XRFC-MO and tracing lacZ mRNA were injected into one cell of four-cell stage embryos, the expression of neural crest makers were examined at stages 157. (A) Knockdown of XRFC by morphoino inhibits the expression of Zic1, Snail2 and FoxD3 in the injected sides (labeled red by staining of the tracing lacZ). (D) Co-injection of XRFC mRNA (800 pg) rescues the expression of Zic1, Snail2 and FoxD3. (G) Overexpression of XRFC expands the expression domains of Zic1, Snail2 and FoxD3. (J, K) XRFC morpholino injection does not affect the expression of Msx1 and Pax3. (L) Twist1 staining of neural crest cells at stage 32 of embryos injected on one side with XRFC-MO or XRFC-MO plus rescue mRNA. In XRFC-MO injected side, the neural crest cells are retained in a dorsal position (arrowhead in L). (P) Quantification of the effect of XRFC-MO on the expression of neural crest markers and the rescue by RFC mRNA. (Q) RT-PCR analysis in animal cap assay to show the effect of XRFC-MO on the expression of the indicated neural crest genes induced by co-injection of tBR and XWnt7b. (R) Transplantation experiment to show the effect of XRFC-MO on neural crest migration. (R) Schematic drawing showing the transplantation procedure. (S) The EGFP-labeled XRFC-MO injected neural crest tissue fails to migrate at tailbud stage after transplantation. (T) Co-injection of XRFC mRNA restores the migration ability of the neural crest cells. inj. side, injected side; con. side, control side. |
