Supplementary Figure 3: The in vitro cytotoxicity and BrdU proliferation assay of GAz4 and CLP reagents in 4T1-luc, C2C12-luc and HepG2-luc cells.
From: Bioluminescent-based imaging and quantification of glucose uptake in vivo
Three different types of cells were incubated for 1 or 24 h with 1 nM-5 mM concentrations of either GAz4 or CLP (upper graph in every panel). Cytotoxicity was determined using AlamarBlue assay as described in the instruction of Invitrogen commercial kit (Invitrogen, DAL1025). Resazurin, the component of this assay, oxidizes NADH to give a highly fluorescent compound and provides information on cellular redox state. Data are presented as mean ± SD (n = 3, biologically independent experiment). Cell viability was calculated as a percentage of viable cells, data were normalized to DMF concentration. Cell proliferation was measured with bromodeoxyuridine (BrdU) assay (bottom graph in every panel). Cells were incubated in growing medium supplemented with BrdU and either GAz4 (100 μM) or CLP (10 μM) for different time intervals (24, 48, and 72 h) at 37 °C and 5% CO2. BrdU incorporation into replicating DNA was measured by commercially available ELISA kit (Abcam, ab126572). According to manufacturer’s instruction. Data are presented as mean ± SD, n = 3, biologically independent experiments.
