FIGURE 2

Spectrum of TFG mutations in human and characterization of Tfg. Linear map of the reported disease‐related mutations in TFG protein. (B) Conservation analysis of the substituted amino acid at 265,269 and 285 in TFG peptides. (C) Multiple‐sequence alignment of the TFG protein sequence in human, mouse, and zebrafish. (D) In situ hybridization histochemistry staining of coronal sections from adult mouse motor cortex and spinal cord. Tfg mRNA was labeled with specific probe (green) and neurons were labeled with NeuN antibody (red), nuclei were labeled with DAPI (blue). Scale bars: 200 μm, 100 μm, or 50 μm. (E) Immunofluorescence staining of primary cultured neurons at day‐8 in culture. The neurites were labeled with MAP2 (green) and the nucleus with DAPI (blue). White arrowhead shows TFG expression in the proximal dendrites of neurons. Scale bars: 50 μm. (F) qRT‐PCR for six embryo development stages (6, 24, 72, 96, 120 and 144 hpf) of wild‐type zebrafish embryos. Hpf, hours postfertilization. (G) Hybridization in situ of tfg in the whole zebrafish embryos at 24, 48 and 72 hpf. The arrow indicates the positive signal in the brain and eye region; the asterisk shows the positive signal in the spinal cord and notochord; the arrowhead shows the positive staining in the pectoral fin anlage.