2.8.1. PMP70 staining

Melanoma cells (10 × 10³) expressing either ASAH1 or non-specific shRNA were plated in multi-well chambered slides. After 24 h, the cells were washed with PBS and fixed with 3.7% paraformaldehyde. The cells were permeabilized using 0.3% Triton X-100. After washing with PBS, the slides were blocked using 5% BSA in PBS. The cells were then probed with PMP70 primary antibody (1:200) (see Supplementary Table 1). After washing, the cells were incubated with HRP-conjugated secondary AlexaFluor-488 anti-rabbit antibody (1:1000) (see Supplementary Table 1). Fluorescence images were collected using a LEICA SP5 confocal laser scanning microscope.