Immunofluorescence Staining of Cell Lines

Fibroblasts and Neuro2A cells were treated as previously described.^{28 (link)} Briefly, cells were fixed with paraformaldehyde at 4% for 30 min at 37°C and then permeabilized using a solution of 10% Triton X-100 diluted 1:5000 in PBS. After 1 h of blocking, cells were incubated with the primary antibody for 40 min at room temperature and, after washes, with the secondary antibody for 20 min at room temperature. Coverslips were mounted using Mowiol solution. Images were acquired using Zeiss LSM700 confocal laser scanning microscopy (Zeiss, Oberkochen, Germany) or the EVOS FL Auto Cell Imaging System (Thermo Fisher Scientific, Waltham, MA, USA) and cells with aggregates were manually counted.