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

Fig. 3. TRAIL-R2/DR5 upregulation is dispensable for sensitization to TRAIL upon glutamine deprivation.

Fig. 3

(A) MDA-MB468 cells were either non-transfected (NT) or transfected with siRNA of GCN2, ATF4, CHOP or Scrambled oligonucleotide (SCR) for 30 h and then incubated with or without glutamine for 24 h. TRAIL-R2 levels determined by RT-qPCR, western blotting and flow-cytometry as described in the Materials and Methods section. (B) MDA-MB468 cells were transfected with siRNA of GCN2, ATF4, CHOP or Scrambled (SCR) for 30 h and then incubated with or without glutamine for 24 h prior to treatment for 24 h in the presence or absence of TRAIL (100 ng/ml). Apoptosis was measured as described in Materials and Methods section. GCN2, ATF4 and CHOP levels were assessed by western blotting. GAPDH was used as a protein loading control. In A (left panel) and B, error bars represent s.d. from three independent experiments. *P < 0.05, n.s. not statistically significant