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. 2014 Apr 24;9(4):e96035. doi: 10.1371/journal.pone.0096035

Figure 4. Inhibition of PI3K/AKT signaling prevents Nrf2-induced depigmentation.

Figure 4

(A) NHEMs were transduced with the indicated adenoviruses (10 MOI) for 6 h. Cells were replenished with fresh medium containing vehicle (DMSO) or the PI3K inhibitor wortmannin (Wort, 100 nM). Cells were cultured for a 2 days, then cells were refed with fresh medium containing wortmannin again. One day after, cellular extracts were prepared and then assessed by Western blotting. Phosphorylation of Akt, mTOR and S6 ribosomal protein by Nrf2 was inhibited by PI3K inhibitor treatment. (B) Tyrosinase (TYR) activity was determined and expressed as a percentage of the control. Data are the means ± SD of triplicate measurements (*P<0.01 vs. control). Nrf2-repressed tyrosinase activity was reversed by PI3K inhibitor treatment. (C) Repression of TYR and TRP1 by Nrf2 was reversed by PI3K inhibitor treatment. (D) The TYR promoter-luciferase reporter adenovirus was co-transduced with the indicated adenoviruses. TYR promoter activity was expressed as a percentage of the control ± SD (*P<0.01 vs. control). Nrf2-repressed tyrosinase promoter activity was reversed by PI3K inhibitor treatment.