Pcmbs

Rat, human, and mouse glioma cell lines F98, U87, U251, and GL261 were obtained from ATCC/LGC-2397 (Wesel, Germany). All cell lines were cultured under standard humified conditions (37°C, 5% CO₂) with DMEM (Biochrom, Berlin, Germany) supplemented with 10% FBS (Biochrom, Berlin, Germany), 1% penicillin/streptomycin (Biochrom, Berlin, Germany), and 1% GlutaMAX (Gibco, Darmstadt, Germany). For detachment, cells were treated with 0.05% trypsin (Biochrom, Berlin, Germany). HUVEC were cultured under standard humified conditions with Endothelial Cell Basal Medium with Endothelial Cell Growth Medium supplement mix (Promo Cell, Heidelberg, Germany). For detachment, cells were treated with StemPro® Accutase® (Gibco, Darmstadt, Germany) according to the manufacturer's instructions. All cells were evaluated under an inverted phase/fluorescence microscope (Olympus IX71, Hamburg, Germany) in the transmitted light path. Image acquisition was achieved via the Cell^F software (Olympus, Hamburg, Germany). To induce hypoxia, cells were treated with 200 μM DFO (Sigma Aldrich, Taufkirchen, Germany). For MCT4 inhibition, cells were treated with 100 μM pCMBS or 150 μM phloretin (both from Sigma Aldrich, Taufkirchen, Germany). Cells were counted using the Cell Counter Z2 (Beckman Coulter, Brea, California, USA) according to the manufacturer's instructions.