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. 2012 Jan 3;120(4):583–589. doi: 10.1289/ehp.1104580

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Stable knockdown of NRF2 and KEAP1 by lentiviral shRNAs in HaCaT cells. (A and C) Silencing effect of NRF2 (A) and KEAP1 (C) in HaCaT cells transduced with shRNA lentivirus targeted against human NRF2, KEAP1, or scrambled (SCR) nontarget negative control (sh-SCR) (n = 3). sh-NRF2-1 through sh‑NRF2-5 indicate shRNAs targeting NRF2; sh-KEAP1-1 through sh-KEAP1-5, shRNAs targeting KEAP1. (B and D) Protein expression of NRF2 (B), KEAP1 (D), and NRF2 target genes NQO1 and GCLC in NRF2-KD and KEAP1-KD cells. Cells were treated with 50 μM tBHQ for the indicated time, and whole-cell lysates were separated on 4–12% Tris-glycine gels. β-ACTIN was used as loading control. (E) Intracellular GSH content in NRF2-KD and KEAP1-KD cells (n = 6). *p < 0.05 versus SCR (A, C, E).