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. 2021 Apr 15;148(7):dev193565. doi: 10.1242/dev.193565

Fig. 6.

Fig. 6.

RIA retrovirus-mediated knockout of Sox10 in neural crest derivatives. (A) Schematic for cloning the Sox10-RIA-CRISPR virus plasmid. (B) Sox10-RIA-CRISPR retrovirus was injected into the neural tube lumen at HH10. At HH25, embryos were fixed, cryosectioned and immunostained. (C) Knockout of Sox10 following emigration from the neural tube does not affect survival but rather causes downregulation of Snail2, a neural crest specification marker. (D-E″) Early (D,D′) and late (E-E″) migrating neural crest cells labeled with Citrine (D′,E″) do not express Snail2 (arrowheads). (F-G″) Transverse section through an injected embryo (F) shows labeled migrating trunk neural crest cells mutant for Sox10 expression close to the sympathetic ganglion (G,G′), with DAPI-labeled nuclei (G″). (H) Labeled cells were also observed in the dorsal root ganglion. (I-I″) Although most Citrine-positive cells were Sox10-negative (unfilled arrowheads), Sox10 protein was detected in a couple of labeled cells (filled arrowheads). drg, dorsal root ganglion; ic, internal carotid; nc, notochord; nt, neural tube; p, pharynx. Dashed line delineates the neural tube in D-E″, and the boundary of the dorsal root ganglion in I-I″.