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. 2022 Jan 10;13:85. doi: 10.1038/s41467-021-27688-5

Fig. 2. Generation of seeded in vitro fibrils.

Fig. 2

(a) Time-resolved ThT fibrillation kinetics of recombinant SAA1.1 protein without (blue) and with 5% (w/w) ex vivo fibrils as seeds (green). The lag phase of the unseeded reaction is 10.9 ± 1.7 h. (b) Cryo-EM image of the seeded in vitro fibrils, showing only fibrils of the major morphology. Scale bar: 100 nm. The micrograph is representative for 1,762 micrographs. (c) Cryo-EM based pitch and width measurements with the major (deep green, superimposing with the orange data points) and the minor (light green) fibril morphology of the seeded in vitro fibrils (n = 20). Orange: ex vivo fibril morphology I; light blue: unseeded in vitro fibril morphology i. The values of pitch/width are: 151.8 ± 2.5 nm/11.9 ± 0.4 nm (ex vivo I); 151.6 ± 2.5 nm/11.9 ± 0.3 nm (major seeded in vitro); 98.9 ± 1.4 nm/8.2 ± 0.4 nm (minor seeded in vitro); 80.1 ± 3.0 nm/9.3 ± 0.4 nm (unseeded in vitro).