fscn1a cooperates with cxcr4a/cxcl12b signaling to regulate directional cranial NC migration. (A) Representative confocal images of cranial NC cells from a 26 hpf Tg(sox10:rfpmb) embryo injected with cxcr4a-GFP mRNA. Arrows highlight Cxcr4a-GFP localized to NC filopodia. Scale bar = 10 µm. (B) Lateral view of leading edge of cranial NC stream 1 in 26 hpf uninjected, cxcl12b mRNA-injected or cxcr4aMO-injected Tg(sox10:rfpmb) or Tg(sox10:rfpmb); fscn1a MZ embryos. Scale bar = 25 µm. (C) Quantitation of filopodia number and filopodia length in uninjected, cxcl12b mRNA-injected or cxcr4aMO-injected Tg(sox10:rfpmb) and Tg(sox10:rfpmb); fscn1a MZ embryos (n = 7 Tg(sox10:rfpmb), n = 9 Tg(sox10:rfpmb) + cxcl12b mRNA, n = 5 Tg(sox10:rfpmb) + cxcr4aMO, n = 7 Tg(sox10:rfpmb); fscn1a MZ, n = 8 Tg(sox10:rfpmb); fscn1a MZ + cxcl12b mRNA, n = 3 Tg(sox10:rfpmb); fscn1a MZ + cxcr4aMO, ***p<0.001, ****p<0.0001). (D) Lateral views of 5 dpf uninjected, cxcl12b mRNA-injected or cxcr4aMO-injected wild type (wt) and fscn1a MZ embryos. Arrowheads indicate reduced jaw. (E) Quantitation of percentage of embryos with normal or abnormal craniofacial development at 5 dpf (n = 43 wt, 104 wt + cxcl12b mRNA, n = 51 wt + cxcr4aMO, n = 165 fscn1a MZ, n = 130 fscn1a MZ + mmMO, n = 123 fscn1a MZ + cxcl12b mRNA, n = 154 fscn1a MZ + cxcr4aMO).
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