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. Author manuscript; available in PMC: 2021 Dec 3.
Published in final edited form as: Mol Cell. 2020 Oct 30;80(5):828–844.e6. doi: 10.1016/j.molcel.2020.10.010

Figure 6. Downregulation of NRF2 Induces Global Enrichment of Selenoproteins Including GPX4.

Figure 6.

(A and B) Immunoblot analysis of NRF2 and GPX4 in the indicated cells. In (B), the blots for NRF2 and β-actin in 3D are from Figure 5L as these experiments were performed concurrently as the experiments presented in Figure 6B. (C) Schematic of proteomics approach. For H1437 cells with shControl, two samples were used for the experiment because the plex was limited to 11 for each TMT experiment. (D) All selenoproteins or proteins required for selenoprotein biosynthesis detected by proteomics in A549 cells transduced with shControl or shNRF2-#2. Data presented are mean log2 (fold change) of shNRF2-#2 compared to shControl cells from three technical replicates. (E) Intensity of all selenoproteins detected in the indicated A549 cells. Data presented are mean ± SD from three technical replicates. Uncorrected Student’s t-test was used to determine statistical significance. *p < 0.05, **p < 0.01, and ***p < 0.001. Unless otherwise noted, the cells were cultured for 72 hours upon treatment with 1 μg/ml doxycycline.