Fig. 7: Analysis of hTTR(59–127) amyloid fibril formation.

RP-HPLC purified fragment was lyophilized and dissolved (0.23 mg/ml) at 25 °C in TBS, pH 7.4. Amyloid fibril formation was tested by ThT binding assay and TEM soon after solubilization of the fragment in TBS. a ThT fluorescence emission in the presence (continuous line) or absence (dashed line) of hTTR(59–127). Measurements were carried out after 1-h incubation of ThT (20 µM) with hTTR(59-127) (50 μg/ml) and sample excitation at 450 nm. Inset: Time-dependent relative fluorescence emission (F/F₀) of ThT (white circles) in the presence of hTTR(59–127), where F and F₀ is the fluorescence intensity recorded after incubation with hTTR(59–127), as indicated. b Representative TEM micrograph of hTTR(59–127) solution. TEM analysis was carried out immediately after solubilization of lyophilized hTTR(59–127) at 0.6 mg/ml concentration. The average length and diameter of hTTR(59–127) fibrils was estimated as 200 ± 50 nm and 20 ± 5 nm, respectively. The scale bar (100 nm) is indicated.