Rabbit anti lamp2a

For immunocytochemistry, SH-SY5Y cells were plated onto 12 mm coverslips (Marienfeld, Germany) as previously described (30) (link). SH-SY5Y cells were transfected with TREM2 construct or mutant plasmids using Lipofectamine 2000. Twenty-four hours after transfection, cells were washed with PBS (140 mM NaCl, 10 mM Na₂HPO₄, 1.75 mM KH₂PO₄ in dH₂O, pH7.4). Cells were fixed in 4% paraformaldehyde for 10 min. After three washes in PBS, cells were permeabilized with PBS containing 0.1% Triton X-100 for 5 min at room temperature. Cells were washed three times and blocked with PBS containing 5% bovine serum albumin for 30 min at 37℃. Cells were then washed three times and incubated overnight with rat anti-HA (1:100; Roche, Switzerland) or rabbit anti-LAMP2A (1:100; Abcam, UK) primary antibodies. After washing five times with PBS, cells were incubated with a secondary antibody coupled to TexasRed (Invitrogen, USA) or Alexa Fluor^Ⓡ 488 (Invitrogen, USA) for 60 min at 37℃. Cells were then washed five times and mounted for imaging. Images were acquired using Zeiss Axiovert 200 LSM510 confocal microscope workstation equipped with a 63x (numerical aperture, 1.4) objective.