Glutamate detection assay kit

The relative levels of glutamate in NE2, A549, stable RBM4-overexpressing or knockdown ESCC cells treated with or without deprivation of glutamine were detected using a commercially available colorimetric determination kit (Glutamate Detection Assay kit, ab83389, Abcam, Cambridge, UK) following the manufacturer’s instructions. For rescue experiments, an equal number of stable cells were seeded in the culture dish and culturing in the medium with DMSO or AMPK inhibitor (5 µM). After 36 h, the decrease of glutamine was examined. For rescue experiments, RBM4 stable knockdown cells were treated with AMPK inhibitor (5 µM) or DMSO (vehicle) at 24 h after seeding an equal number in 10 cm dishes. After 36 h, the levels of intracellular glutamate were examined.