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. 2022 May 27;2022:9626703. doi: 10.1155/2022/9626703

Figure 4.

Figure 4

PTS binds to the Kelch domain of Keap1 to regulate Nrf2 activity. (a) Representative Western blotting analysis of Nrf-2 in the cytoplasm and nuclear fractions as well as Keap1 expression in whole-cell lysates. (b–d) Nuclear Nrf2, cytoplasm Nrf2, and Keap1 levels were quantified using ImageJ software. Data are expressed as the mean ± SEM (n = 3). ∗P < 0.05, ∗∗P < 0.01; ns: not significant. (e) In silico molecular docking of PTS with Keap1. A two-dimensional diagram shows the interactions of PTS to the amino acid residues in the binding pocket of Keap1. (f) Anti-Keap1 immunoprecipitation was analyzed by immunoblot with anti-Nrf2 antibodies for detection of Keap1-conjugated Nrf2. (g) Anti-Nrf2 immunoprecipitation was analyzed by immunoblot with anti-ubiquitin antibodies for detection of ubiquitin-conjugated Nrf2. Data are expressed as the mean ± SEM (n = 3). ∗P < 0.05, ∗∗P < 0.01; ns: not significant.