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Fig. 2 | Molecular Brain

Fig. 2

From: Knockdown of Son, a mouse homologue of the ZTTK syndrome gene, causes neuronal migration defects and dendritic spine abnormalities

Fig. 2

SON is necessary for normal neuronal migration. a A schematic representation of the shRNA target sites. The closed and open arrowheads indicate the target positions of shRNA#1 and shRNA#2, respectively, on SON mRNA. b Characterization of Son shRNA-expressing vectors. Neuro-2a cells were transfected with parental pLLC vector (control) or vectors engineered to express Son shRNAs (shRNA#1 or shRNA#2). Cell lysates were subjected to Western blotting with an anti-SON antibody to assess the effect of knockdown. β-actin was used as a loading control and was used to normalize quantified values of SON signals. c Representative images showing the distribution of GFP-positive cells transfected with either empty vectors (control) or a vector expressing shRNA#1 or shRNA#2 in the developing mouse cerebral cortex. The layered structure is shown on the left. UCP: upper cortical plate; LCP: lower cortical plate; other abbreviations as described in Fig. 1d. The cortical plate is divided into the UCP and LCP according to cell density [18]. Transfection was performed by IUE at E14.5. Coronal brain sections were prepared at E18.5 and stained for GFP (green) and DAPI (blue). d The quantification of GFP-positive cells in each layer of the developing cortex. Each layer is described as in (c). Total numbers of GFP-positive cells studied in each brain ranged 306–567. Error bars represent standard error of the mean (SEM). n = 4; *p < 0.05 by one-way ANOVA followed by Dunnett’s test. The original data are available in Additional file 1 [Table S1]. e The confirmation of Son knockdown in shRNA-introduced neurons. Coronal sections prepared as described in (c) were stained for GFP (green), SON (red), and DAPI (blue)

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