FIGURE 3 RAW DATA MYD88 WT FULL LENGTH DILUTIONS SEEDED

Number of photons from time traces from approx 50 MYD88 dilutions with and without MYD88 cherry tagged seeds, raw data analysed in additional files corresponding to (A) Schematic diagram of the principle of two-colour seeding experiments testing the self-replication propensity of full-length MyD88 filaments. Full-length MyD88is expressed in an mCherry-tagged version above its supercritical concentration to create filaments, which are gently spun and washed, then sonicated to increase the number of fragments. These “seeds” are then mixed in a sample expressing GFP-tagged full-length MyD88 at sub-critical concentrations. (B) Example of fluorescence time trace for MyD88 at 10 nM concentration. Unseeded sample demonstrating a monomeric time trace profile (above) with the seeded sample (below) showing polymerisation of GFP-MyD88 upon the addition of MyD88 ‘seeds’. (C) The B parameter (brightness) correlating with number of oligomers detected in typical time-traces as a function of protein concentration (nM), with and without “seeds” introduced. The subcritical, supercritical and “meta-stable” zones are labelled. Values are from approx. 50 repeated dilution experiments with the various protein concentrations and corresponding brightness values obtained plotted.

本数据集包含约50组分别带有与不带有mCherry标记（mCherry）的MYD88种子的MYD88稀释样本的时间轨迹光子计数数据，原始数据分析结果收录于对应以下(A)至(C)内容的附加文件中，具体如下：
(A) 双色种子实验原理示意图：该实验用于检测全长MYD88丝状体的自我复制倾向。将带有mCherry标记的全长MYD88在超临界浓度下表达以形成丝状体，经温和离心洗涤后通过超声破碎增加片段数量，随后将这些“种子”与亚临界浓度下表达GFP标记（GFP，Green Fluorescent Protein）的全长MYD88样本混合。
(B) 10 nM浓度下MYD88的荧光时间轨迹示例：未添加种子的样本呈现单体时间轨迹特征（上图），添加种子的样本（下图）则显示GFP标记的MYD88在加入MYD88“种子”后发生聚合反应。
(C) 与典型时间轨迹中检测到的寡聚体数量相关的B参数（亮度）随蛋白质浓度（nM）的变化关系，分别对应添加与不添加“种子”的实验组。图中标注了亚临界、超临界及“亚稳态”区域，所有数据来自约50组重复稀释实验，涵盖不同蛋白质浓度下获得的对应亮度值并完成绘图。