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. 2020 Mar 1;12(3):569. doi: 10.3390/cancers12030569

Figure 2.

Figure 2

NRF2 activation by PERK partially relies on ROS generation during protein synthesis recovery. NCI-H358 cells were treated with AP over a time course of 24 h and RT-qPCR analysis were performed on canonical (a) ATF4-target genes (TRB3, GADD34, CHOP) or (b) NRF2-target genes (NQO1, HO1, GCLC). (c) Time course analysis of the KEAP1/NRF2 complex dissociation following AP treatment. CI-H358 cells were transiently co-transfected with the Neh2-Luc reporter and Renilla vectors and treated with NAC for 1 h prior to the addition of AP for the indicated periods of time. Data were normalized against the Renilla luciferase activity for each condition. Activity fold increase is reported. (d) Western blot analysis of ATF4, NRF2 and CHOP in cytoplasmic and nuclear fractions upon AP-mediated Fv2E-PERK activation with or without NAC. * = non-specific bands. (e) RT-qPCR analysis of NQO1, HO1 and GCLC expression levels. NCI-H358 cells were treated with or without NAC for 1 h prior to the addition of AP for 24 h. (f) RT-qPCR analysis of NQO1, HO1 and GCLC expression levels. NCI-H358 cells were treated with or without NAC for 1 h prior to the addition of tunicamycin for 24 h. Data are expressed as means ± SEM of at least three independent experiments, * p < 0.05, ** p < 0.01, *** p < 0.001.