FIGURE 2 RAW DATA

(A) Increase of apparent brightnesses of diffusing species. Values of B parameters obtained for expressions at protein concentrations >150 nM were averaged and normalised to the sfGFP monomer control. Clearly, the data show that only full-length MyD88 (red) is capable of forming large complexes, while the TIR domain (green) or the death domain (blue) form smaller oligomers. Inset: Expansion of the values obtained for TIR and death domains compared to sfGFP control. (B) FCS data in solution. Control based on GFP monomer (black). Correlation curves obtained for the TIR domain (green), death domain (blue) and full-length MyD88 (red). A clear shift in diffusion time can be seen between the full-length protein and the separate domains. FCS curves are representative traces from 3 repeated measurements. (C) Hydrodynamic radius normalised by the sfGFP control (black) calculated for the domains and full-length MyD88, indicating the increase of approximate physical size of the oligomeric species. Values are mean ± SD from 3 repeat measurements.

(A) 扩散粒子的表观亮度升高。针对蛋白浓度＞150 nM时的拟合表达式所得到的B参数取平均值，并以超折叠绿色荧光蛋白单体（superfolder GFP, sfGFP）对照进行归一化处理。显然，实验数据表明仅全长髓系细胞分化因子88（Myeloid differentiation primary response 88, MyD88，红色曲线）能够形成大型复合物，而Toll/白介素-1受体结构域（Toll/interleukin-1 receptor domain, TIR domain，绿色曲线）与死亡结构域（death domain，蓝色曲线）仅可形成小型寡聚体。插图：相较于sfGFP对照，TIR结构域与死亡结构域的数值范围被放大展示。
(B) 溶液中的荧光相关光谱（Fluorescence Correlation Spectroscopy, FCS）数据。以GFP单体为对照（黑色曲线）。分别得到TIR结构域（绿色曲线）、死亡结构域（蓝色曲线）与全长MyD88（红色曲线）的相关曲线。可观察到全长蛋白与单独结构域的扩散时间存在显著偏移。本次展示的FCS曲线为3次重复测量的代表性轨迹。
(C) 针对各结构域与全长MyD88计算得到的、以sfGFP对照归一化的流体力学半径（黑色曲线），结果表明寡聚体物种的近似物理尺寸有所增加。数值为3次重复测量的平均值±标准差（standard deviation, SD）。