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. 2018 Oct 3;293(47):18242–18269. doi: 10.1074/jbc.RA117.001245

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© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license.

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Figure 6. — Mini-GAGR reduces the colocalization and interaction between Keap1 and Nrf2. A and B, mouse cortical neurons (E17, DIV 14) were treated with 1 μm mini-GAGR (mini) for 0 h (A) and 3 h (B) and processed for immunocytochemistry with primary antibodies against Nrf2 (green) and Keap1 (red) and secondary antibodies (Alexa Fluor 488 and Alexa Fluor 594), Pearson's colocalization coefficient R was quantified using ImageJ. Scale bar, 30 μm. C, mouse cortical neurons (E17, DIV 14) were treated with either vehicle (control) or 1 μm mini-GAGR (mini) for 3 h and lysed to obtain cytosols for co-IP using either control IgG or anti-Nrf2 IgG (αNrf2). The band densities of precipitated Nrf2 (80 kDa) and Keap1 (70 kDa) proteins were quantified by ImageJ to obtain average density ± S.E. (error bars) for the bar graphs. D, Nrf2, control versus mini-GAGR. E, Keap1, control versus mini-GAGR (n = 3 different embryo batches; *, p < 0.05; Student's t test, two-tailed). Data are expressed as mean ± S.E.