Fig. 4.

Overexpression of Fbxo9 reduces Neurog2 protein expression and directs NCCs to differentiate into glial lineage. (A) Immunofluorescence for Neurog2 on transverse sections of embryos treated with GFP (n = 6), Sox10 (n = 6), and Fbxo9 (n = 7) at 24 hpt (or HH16). (a′, a″, and a″′) The magnified areas are marked with dashed boxes. Nuclei were counterstained with DAPI. Solid white and open arrowheads indicate cells with nuclear Neurog2 and nuclear+cytoplasmic Neurog2 expression, respectively. (B) Graph showing ratio of Neurog2⁺ cells between electroporated and unelectroporated sides of neural tubes treated with the indicated constructs. Quantification of the number of GFP⁺Neurog2⁺ cells in embryos treated with the indicated constructs. Graph showing the percentage of cells with nuclear Neurog2 only and nuclear+cytoplasmic Neurog2 in the total number of GFP⁺ cells from embryos treated with the indicated constructs. (C) Whole-mount immunofluorescence for GFP, HuC/D, and Sox2 on embryos treated with GFP (n = 6) or Fbxo9 (n = 7) at 48 hpt. White dotted lines indicate the plane of sectioning. (D) Quantification of the number of cells expressing either GFP⁺ alone, GFP⁺HuC/D⁺, or GFP⁺Sox2⁺ in embryos treated with the indicated constructs. (E) Quantification of the number of HuC/D⁺ cells in the electroporated side of embryos treated with the indicated constructs. (F) In situ hybridization for Fabp7 expression on cross-sections of embryos treated with the indicated constructs at 48 hpt followed by immunofluorescence for GFP. The magnified areas are marked with dashes boxes. Black arrowheads indicate cells coexpressing GFP and Fabp7. Error bars ± SEM. ns, nonsignificant (***P < 0.001). (Scale bars: embryos, 10 μm; sections, 50 μm.)