Vesicle trafficking analysis

SH-SY5Y cells were transduced with pWPI vectors expressing the T30 and CW-T30 isoforms and then, were fixed and stained with antibodies against β-tubulin (SIGMA, T4026), (GFP (ABCAM, 13970), Tau (NUVUSBIO, NB100-822247), LC3 (SIGMA, L7543), EEA1 (BD Biosciences, 610457), Rab11A1 (Invitrogen, 71-5300), CD63 (DSHB, H5C6), LAMP1 (DSHB, H4A3), and with DAPI (4',6-diamidino-2-phenylindole, Merck, 268298). When indicated, cells were treated with LysotrackerRed DND-99 (Invitrogen, L7528) before fixing. Confocal microscopy analysis was performed using Fiji software and vesicles were segmented using CellPose Plug-in.