Figure 7.

GPC3-Unc5 signaling determines neuroblastoma cell migration properties

(A) UMAP visualization of single-cell data from neuroblastoma tumors (Dong et al., 2020) and quantification for selected transcripts.

(B) Heatmap of Unc5A-D and GPC3 mRNA expression in 4 human neuroblastoma cell lines, measured by qRT-PCR.

(C) SY5Y:GFP cells were transfected (scramble, scr., or GPC3 siRNA) and engrafted within the migratory trunk neural crest of E2 chicken embryos and slices analyzed 2 days later. Neural crest-derived structures were labeled with an anti-HNK1 antibody, nuclei with Hoechst.

(D) Quantification of SY5Y cell and tumor positions two days after grafting.

(E and F) As (C) and (D), but SY5Y cells were transfected with vectors encoding rUnc5D^IgIgTSP, Unc5D^IgIgTSPGU or pCAGIG (control) vectors prior to the graft. Samples were labeled with anti-human mitochondrial antibody to reveal all SY5Y cells (transfected: green + red; non-transfected: red).

(G and H) SY5Y cells were transfected with vectors encoding Nano^glue or Nano^break prior to the graft. Scale bars: 200 μm.

For (D), (F), and (H), we used χ² tests to compare scr. (control) versus GPC3 siRNA conditions (D), rUnc5d^ecto versus rUnc5d^ectoGU (F), and Nano^glue versus Nano^break conditions (H).

NT: Neural Tube; S: Somite; DRG: Dorsal Root Ganglia; SG: Sympathetic Ganglia; AG: Adrenal Gland; DA: Dorsal Aorta; Me: Mesonephros.

See also Figure S7.