Figure 3.

TRPML-1 nuclear localization in glioblastoma cell lines. (a) Proteins derived from membrane fraction (Mem), cytosolic fraction (Cyto), nuclear/cytoskeletal fraction (Nuc), and whole cell lysate (WCL) were immunoblotted with anti-TRPML-1 Ab. Whole cell lysate was used as control. The purity of subcellular fractions was assessed by blotting against specific markers. Cytosolic and membrane marker: GAPDH; membrane-bound organelles markers: LAMP-1; nuclear marker: Histone H3. Blots are representative of one of three separate experiments. (b) To analyze the ability of TRPML-1 to bind DNA, nuclear fraction (Nuc) proteins and DNA were isolated from T98 and U251. The samples were electrophoresed in SDS-PAGE gel and incubated with anti-TRPML-1 Ab to determine the relative protein expression. Data are representative of three separate experiments.