*2.14. Cell Fractionation*

Cells were washed with PBS and lysed at a concentration of 1 × 10<sup>4</sup> cells per μl in protein separation bu ffer A (0.2% Triton X-100, 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 2.5 mM EDTA) [32]. As misfolded proteins aggregate and become resistant to Triton X-100 solubilisation [32,35], Triton X-100-insoluble fraction was separated by centrifugation at 3400 *g* for 10 min. Pellets were enriched in misfolded proteins. Samples were analysed by immunoblot. Loading was normalised by the number of lysed cells. Band intensities were determined with ImageJ software (version 1.50i, NIH, Washington, WA, USA, 2016) and soluble/insoluble ratios calculated.
