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. 2015 Feb 10;172(7):1869–1881. doi: 10.1111/bph.13031

Figure 5.

Figure 5

UFP-512-induced Nrf2 translocation from the cytoplasm to nucleus after H/R. (A) Representative Western blots of nuclear Nrf2 protein after DOR activation and/or inhibition in normoxia and H/R. (B) Relative quantitation of Nrf2 protein in the nucleus. (C) Relative quantitation of Nrf2 protein in the cytoplasm. After treatment with UFP-512 and/or naltrindole during the H/R procedure, nuclear and cytoplasmic fractions of the HEK293t cells were extracted for Western blotting and Nrf2 determined with anti-Nrf2 antibody. C, normoxic control. —, H/R alone. U, DOR activation with UFP-512. N, DOR inhibition with naltrindole. U + N, UFP-512 and naltrindole co-administration.*P < 0.05, **P < 0.01, ***P < 0.005, significantly different from control in normoxia, #P < 0.05, ##P < 0.01, significantly different from H/R alone. ◊P < 0.05, significantly different from UFP-512 after H/R stress. At least three independent experiments were performed. Note that H/R stress decreased cytoplasmic Nrf2 protein and increased nuclear Nrf2 protein, suggesting a translocation from the cytoplasm to nucleus. DOR activation with UFP-512 further enhanced such translocation, whereas DOR antagonism with naltrindole abolished the UFP-512-enhanced Nrf2 translocation.