IF analysis was performed as described from previous reports [5 (link), 7 (link)]. Cells were fixed in 10% trichloroacetic acid and permeabilized with 0.5% Triton X-100 in PBS. After blocking with PBS containing 1% bovine serum albumin and 0.01% Triton X-100 for 1 h at 4°C, the cells were incubated with mouse anti-Cathepsin D (dilution: 1/1000) overnight at 4°C. Bound antibody was visualized using Alexa Fluor 488 anti-mouse IgG antibody (Invitrogen, dilution: 1/2000). The cells were mounted in VECTASHIELD Mounting Medium with DAPI (Vector Laboratories) and observed under a fluorescence microscope (DM6000 B, Leica, Wetzlar, Germany).
Free full text: Click here
