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. 2016 Jul 18;6:30033. doi: 10.1038/srep30033

Figure 3. GSH regulates mTORC1 signaling and the ISR during cystine starvation.

Figure 3

(A) HepG2 cells were treated with or without cystine (100 μM) in the presence or absence of OU749 (250 μm) for 9 h, and GSH (1 mM) was supplemented during the final 3 h for the indicated groups. (B) Cells were treated with or without cystine (100 μM) in the presence or absence of OU749 (250 μM) for 9 h, and GSHee (5 mM) was supplemented during the final 3 h for the indicated groups. Immunoblotting was performed to analyze the phosphorylated and total proteins. (C) Cells were pretreated with or without BSO (300 μM) for 18 h, and then stimulated with or without cystine (100 μM) in the presence or absence of BSO for (300 μM) 9 h. Intracellular GSH was measured with GSH colorimetric detection kit. Data are expressed as means ± SEM of n = 5, #p < 0.01. (D) Cells were pretreated with or without BSO (300 μM) for 18 h, and then stimulated with or without cystine (100 μM) in the presence or absence of BSO (300 μM) for 4 h. Immunoblotting was performed to analyze the phosphorylated and total proteins.