Subcellular distribution of TRPML-1 in glioblastoma cell lines. Cells were fixed, permeabilized, and stained with anti-human TRPML-1 Ab followed by Alexa Fluor-594 secondary Ab. 4′,6-diamidino-2-phenylindole (DAPI) was used to counterstain nuclei. (a) Confocal microscopy analysis of TRPML-1 expression in glioma cells and PBMC, used as positive control. Calibration bar: 20 μm. (b) Z-Stack of glioma cells, stained as described above was performed using confocal microscopy. Pictures were taken on several planes, ranging from upper to lower levels. Calibration bar: 20 μm. (c) Colocalization with endolysosomal compartment was analyzed by staining untransfected and siTRPML-1 transfected cells with anti-LAMP-1 Ab, followed by incubation with Alexa Fluor-488 secondary Ab. Calibration bar: 30 μm.