FIGURE 1 ANALYSED RAW DATA

. For GFP-tagged MyD88 TIRdomain, small fluctuations in intensity are recorded around the average fluorescence value, as expected for a low-order oligomer such as a dimer (for example, if 20 proteins are detected simultaneously, the exit/entry of a single protein causes a decrease/increase of signal of only 5%). The GFP-tagged MyD88 DD shows larger bursts of fluorescence correlating with these death domains forming higher-order oligomeric complexes. As seen by the fluorescent time traces, N-terminally GFP tagged full-length MyD88 shows extremely large filamentous polymers of MyD88 diffusing through the confocal volume. (B) The B parameter (Brightness) correlates with the number of oligomers detected in typical time-traces as a function of protein concentration (nM), for the TIR domain (green), DD (blue) and wild-type full length MyD88 (red). Protein concentrations range from 0 to 320 nM. Monomeric GFP (black) is included as a control. Inset: expansion of the signals obtained for the individual domains, over a lower concentration range. Fluorescence Intensity Time traces in (A) are representative traces obtained at >200nM protein concentrations. Values in (B) are from approx. 30 dilution experiments with the various protein concentrations and corresponding brightness values obtained plotted.