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. 2018 Feb 12;9(2):205. doi: 10.1038/s41419-018-0263-0

Fig. 7. Treatment with l-Asparaginase sensitizes TNBC cells to TRAIL.

Fig. 7

MDA-MB468 cells were incubated in medium with glutamine (+GLN), without glutamine (-GLN) or with glutamine and l-Asparaginase (l-ASNase) for 48 h. Glutamine (2 mM) was added to some cultures for the last 6 h of incubation with l-ASNase (l-ASNase + GLN). (A) ATF4 levels were assessed by western blotting. CHOP and TRAIL-R2 mRNA levels were determined by RT-qPCR. Error bars represent SD from three independent experiments. **P < 0.01. (B) TRAIL-R2 levels were assessed either by western blotting or by flow-cytometry with a TRAIL-R2-PE antibody as described in the Materials and Methods section. Cells incubated with control IgG-PE antibody were used as a control for background fluorescence. (C) FLIP levels were assessed by western blotting. GAPDH was used as a protein loading control. Data are representative of at least three independent experiments (panels B and C). In (D) MDA-MB468 cells incubated as described above were treated for 24 h in the presence or absence of TRAIL (100 ng/ml) and apoptosis was measured as described in Materials and Methods section. Error bars represent s.d. from three independent experiments. ***P < 0.001. n.s. not statistically significant.