Fig. 1: S-palmitoylation is required for maintaining GLUT1 PM localization.
From: DHHC9-mediated GLUT1 S-palmitoylation promotes glioblastoma glycolysis and tumorigenesis
a U87 or T98G cells were treated with DMSO or 50 μM of 2-BP for 8 h followed by incubation with 1 µg ml−1 of cholera toxin subunit B (CTB) conjugated with Alexa Fluor 594 for 5 min at 37 °C. Endogenous GLUT1 cellular localization was visualized by immunofluorescent staining using antibodies against GLUT1. PM plasma membrane. Scale bar, 20 μm. b U87 or T98G cells were treated with DMSO or 50 μM of 2-BP for 8 h. Levels of GLUT1 in PM and ICM fractions were analyzed by immunoblotting. ATP1A1, calnexin, and GAPDH served as the marker of PM, ICM, and cytosol fraction, respectively. PM plasma membrane, ICM intracellular membrane, WCL whole-cell lysate. c GLUT1 palmitoylation was analyzed in lysates derived from U87 or T98G cells metabolically labeled with a palmitoylation probe (50 μM alkynyl palmitic acid [PA]) for 4 h by click reaction and streptavidin bead pulldown in the absence or presence of hydroxylamine (HAM), followed by immunoblotting using indicated antibodies. d GLUT1 palmitoylation levels in U87 or T98G cells were analyzed by the APE assays, upon 50 μM of 2-BP treatment in the absence or presence of HAM. PEG-GLUT1 bands indicated palmitoylated GLUT1. Representative results were obtained from at least three independent experiments with similar results. Source data are provided as a Source Data file.
