Fig. 3
From: Hypertonicity-enforced BCL-2 addiction unleashes the cytotoxic potential of death receptors
Changes in tonicity do not affect TRAIL-receptor expression and receptor-ligand interaction. a Cell surface expression of TRAIL-R1, TRAIL-R2, TRAIL-R3, and TRAIL-R4 was analyzed in HCT116 cells using flow cytometry. Data shown are representative of two experiments performed. b and c HCT116 cells were challenged with the indicated concentrations of b FLAG-TNC-TRAILmutR1 and c FLAG-TNC-TRAILmutR2 (both artificially cross-linked by adding anti-FLAG antibody) in the presence and absence of NaCl (75 mM). d HCT116 cells were treated with the indicated concentrations of the TRAIL-R2 targeting, agonistic antibody conatumumab (artificially cross-linked by adding protein G) in the presence and absence of NaCl (75 mM). For b–d, data points and mean ± SEM from three independent experiments are shown. e and f Equilibrium binding studies were performed by incubating SK-Mel-3 cells with the indicated concentrations of e GpL-FLAG-TNC-TRAIL and f GpL-FLAG-conatumumab as detailed in “Materials and methods.” Binding curves and R2 values shown are representative of three experiments performed; KD values shown were calculated from three independent experiments. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, *p < 0.0001; n.s., not statistically significant; RLU, relative light units
