Figure 6
CarD forms amyloid-like fibrils in solution. (a) Thioflavin T fluorescence assay: Staining of native CarD (5, 10 and 100 μM) and CarDtr (100 µM) with amyloid specific fluorescent dye, Thioflavin T (ThT) at 37 °C after 15 min shows enhanced fluorescence intensity in case of native CarD as compared to CarDtr. The average fluorescence intensity and standard error bars have been calculated from three independent experiments. (b) The kinetics of amyloid-like fibril formation of native CarD (5 μM) in the absence and presence of 5% and 10% seeds monitored at 37 °C. The ThT fluorescence intensity was measured using excitation wavelength of 440 nm and emission wavelength of 482 nm. (c and d) The transmission electron microscopy (TEM) was used to visualize the CarD amyloids. The electron micrographs reveal several branched amyloid-like fibrils (c) and spikes (the aggregates of amyloid-like fibrils; d).
