Fig. 6

Role of Syntaxin1A in neuronal migration during corticogenesis. a Syntaxin1A distribution at E17. Coronal section was double-stained for Syntaxin1A (i and ii) and nuclei (ii). Bar, 100 μm. b Subcellular distribution of Syntaxin1A in migrating neurons in the CP. pCAG-GFP was electroporated into cerebral cortices at E14.5 and fixed at E17. A coronal section was prepared and stained for GFP (i) and Syntaxin1A (ii). Merged image was also shown (iii). Bar, 5 μm. c Characterization of vectors. pCAG-Myc-mSyntaxin1A was transfected into COS7 cells with pSuper-H1.shLuc (Control), sh-Stx1A + pCAG-Myc (vector), or sh-Stx1A + pCAG-Myc-Syntaxin1A-R (i). pCAG-Myc-mSyntaxin1B was transfected with pSuper-H1.shLuc (Control) or sh-Stx1A (ii). After 48 h, cells were harvested and subjected to western blotting (10% gel) with anti-Myc. Anti-Sept11 was used for a loading control. d Knockdown of endogenous Syntaxin1A in cortical neurons. pCAG-GFP was transfected with pSuper-H1.shLuc (Control), or sh-Stx1A into dissociated neurons obtained at E14 and cultured for 48 h. Then, cells were fixed and immunostained for GFP (green) and Syntaxin1A (red). Nuclei were visualized by DAPI. Bar, 10 μm. e Migration defects of Syntaxin1A-deficient cortical neurons and rescue experiments. pCAG-RFP was electroporated in utero with pSuper-H1.shLuc (i), sh-Stx1A (ii), sh-Stx1A + pCAG-Myc (iii) or sh-Stx1A + pCAG-Myc-Syntaxin1A-R (i’) into E14.5 embryonic brains. Coronal sections were prepared at P0 and stained with anti-RFP (red) and DAPI (blue). Bar, 100 μm. f Quantification of the distribution of neurons in bin 1–5 and IZ in (e). Error bars indicate SD (i, n = 4; ii, n = 6; iii, n = 4; i’, n = 5); **p < 0.01 *p < 0.05 by Tukey-Kramer LSD