Fig. S1
Radial glia and neuronal ablation by the NTR/Mtz-mediated cell ablation method. (A–A″) 78 hpf Tg(gfap:GFP);TgBAC(gfap:gal4ff);Tg(UAS:mcherry-NTR) trunk spinal cord section. GFP+ radial glia and mCherry+ cells are largely colocalized. (B–B″) 78 hpf TgBAC(gfap:gal4ff);Tg(UAS:mcherry-NTR) trunk spinal cord section immunostained for HuC/HuD (green). EGFP+ neurons and mCherry+ cells are mostly distinct. (C) Time course of NTR/Mtz-mediated cell ablation of radial glia for D–F″. (D) 78 hpf TgBAC(gfap:gfap-EGFP);TgBAC(gfap:gal4ff);Tg(UAS:mcherry-NTR) trunk. TgBAC(gfap:Gfap-EGFP) expression and TgBAC(gfap:gal4ff);Tg(UAS:mCherry-NTR) expression are observed in the spinal cord in an overlapping manner. SC, spinal cord. (E–E″ and F–F″) 154 hpf TgBAC(gfap:gfap-EGFP);TgBAC(gfap:gal4ff);Tg(UAS:mcherry-NTR) trunk after treatment with DMSO (E–E″) or Mtz (F–F″) starting at 78 hpf. Unlike DMSO-treated fish that show strong coexpression of TgBAC(gfap:Gfap-EGFP) and TgBAC(gfap:gal4ff);Tg(UAS:mCherry-NTR) in their spinal cord, Mtz-treated fish show a dramatic reduction of this coexpression. (G) Time course of NTR/Mtz-mediated cell ablation of neurons for H–J″. (H) 78 hpf Tg(elavl3:EGFP);Tg(elavl3:gal4);Tg(UAS:mcherry-NTR) trunk. Tg(elavl3:EGFP) expression and Tg(elavl3:gal4);Tg(UAS:mCherry-NTR) expression are observed in the spinal cord in an overlapping manner. (I–I″ and J–J″) 154 hpf Tg(elavl3:EGFP);Tg(elavl3:gal4);Tg(UAS:mcherry-NTR) trunk after treatment with DMSO (I–I″) or Mtz (J–J″) starting at 78 hpf. Unlike DMSO-treated fish that show strong coexpression of Tg(elavl3:EGFP) and Tg(elavl3:gal4);Tg(UAS:mCherry-NTR) in their spinal cord, Mtz-treated fish show a dramatic reduction of this coexpression. (Scale bars, 100 µm in D, F″, H, and J″ and 20 µm in A″ and B″.) |