Source Data For Neurite Outgrowth Assay For The: Nrcam Is A Marker For Substrate-Selective Activation Of Adam10 In Alzheimer'S Disease

<strong>Neurite outgrowth assay:</strong> 1x10⁵ neurons were plated into XONA-microfluidic chambers (standard neuron device, SND450) that had been placed on PDL coated coverslips, according to the manufacturers’ instructions. After 4h the plated cells were infected with the respective viruses (scr. shRNA-EGFP control, or NrCAM shRNA-EGFP, 1:1000). Cells were kept until DIV3 at 37°C and 5% CO₂, and then the first photomicrographs of the fluorescent neurons were taken with a Leica DM6000 inverted microscope. The images covered the whole channel area in the middle of the respective chambers. Afterwards, the neurons were treated with GI254023x (5 µM), or vehicle and kept at 37°C and 5% CO₂ for 24h. At DIV4, a second set of photomicrographs of the same areas were taken and analyzed for length differences of single neurites (length in mm at 24h-0h) with Leica LASX software. Only neurites that had already entered and not yet left the channels on the other side at the timepoint 0h were used for the calculation. When neurites were separating after leaving the main channel the longest process was quantified.<strong>神经突生长测定（Neurite outgrowth assay）</strong>：将1×10⁵个神经元接种至预先置于聚赖氨酸（PDL）包被盖玻片上的XONA微流控小室（XONA-microfluidic chambers，型号为标准神经元装置SND450），操作严格遵循制造商说明书。接种4小时后，使用对应病毒感染细胞：分别为scrambled shRNA-EGFP对照病毒（scr. shRNA-EGFP对照）或神经细胞黏附分子（NrCAM）短发夹RNA-EGFP（NrCAM shRNA-EGFP）病毒，感染比例为1:1000。将细胞置于37℃、5% CO₂环境中培养至体外培养第3天（DIV3），随后使用徕卡DM6000（Leica DM6000）倒置显微镜拍摄首批荧光神经元显微图像，图像覆盖对应小室中部的全部通道区域。后续将神经元用GI254023x（5 μM）处理，或仅添加溶剂对照（vehicle），继续在37℃、5% CO₂条件下培养24小时。至体外培养第4天（DIV4），拍摄同一区域的第二批显微图像，并使用徕卡LASX（Leica LASX）软件分析单个神经突的长度差异（以24h与0h时间点的长度差值计，单位为mm）。仅选取在0h时间点已进入通道且尚未从通道另一侧穿出的神经突用于后续计算。若神经突从主通道穿出后发生分支，则以最长的突起作为量化对象。