Downregulation of ADAM10 induces neuronal differentiation in the ventral spinal cord.

After electroporation at E4, transverse sections at E6 were used for immunostaining. The transfected cells are marked with green fluorescence, and the control morpholinos (Fluo-m) and the untransfected side (left side) serve as controls. The immune reactive cells are stained red. Cell nuclei are labeled by DAPI (blue). (A–X) Immunohistochemistry was performed in adjacent sections with antibodies against Evx1 (A–H), En1 (I–L), MNR2 (M–P), Islet1 (Q–T), and Lim3 (U–X), respectively. Arrows indicate immune reactive cells in the prospective ventricular zone and arrowheads indicate no immune reaction in the transfected region (green). Dotted lines indicate the boundary of the alar plate and the basal plate. (Y–B’) Computer-generated overlay of neurons with different domain markers such as En1 (red) and Evx1 (green) in (Y); En1 (red) and MNR2 (green) in (Z); Lim3 (red) and MNR2 (green) in (A’); and MNR2 (red) and Islet1 (green) in (B’). Arrowheads indicate the expanded cells over the boundary into adjacent domains. Dotted lines indicate the boundary of the different domains. (C’) Quantitative data of the cell number in different domains normalized by the number of the control side, which is set to be 1. All data are presented as mean ± SEM from at least 3 independent experiments (*p<0.05, **p<0.01 compared to the control). Scale bar, 200 µm in (A) for (A–X).

在胚胎第4天（E4）实施电穿孔后，于胚胎第6天（E6）取材制备横向切片用于免疫染色实验。转染细胞以绿色荧光标记，对照吗啉寡核苷酸（Fluo-m）与未转染侧（左侧）作为实验对照。免疫反应阳性细胞被染为红色，细胞核由DAPI（蓝色）标记。
(A–X) 分别使用针对Evx1（A–H）、En1（I–L）、MNR2（M–P）、Islet1（Q–T）及Lim3（U–X）的抗体，对相邻切片开展免疫组织化学检测。箭头指向预期脑室区中的免疫反应阳性细胞，箭头头则指示转染区域（绿色荧光标记区域）未出现免疫反应。虚线标示翼板（alar plate）与基板（basal plate）的边界。
(Y–B’) 计算机生成的不同结构域神经元标记叠加图：其中(Y)为En1（红色）与Evx1（绿色）的叠加图；(Z)为En1（红色）与MNR2（绿色）的叠加图；(A’)为Lim3（红色）与MNR2（绿色）的叠加图；(B’)为MNR2（红色）与Islet1（绿色）的叠加图。箭头头指示跨越结构域边界侵入相邻区域的扩增细胞。虚线标示不同结构域的边界。
(C’) 以对照组侧的细胞数设为1，对不同结构域内的细胞数进行归一化处理后的定量数据。所有数据均以至少3次独立实验的平均值±标准误（SEM）呈现（与对照组比较：*p<0.05，**p<0.01）。(A)的标尺为200 µm，适用于(A–X)所有子图。