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. 2008 Jun 27;36(13):4327–4336. doi: 10.1093/nar/gkn416

Figure 2.

Figure 2.

APE1/Ref-1 promotes GSH- or Trx-mediated reduction of p50. (A) The p50 Cys-62 is a target for the synergistic effect of APE1/Ref-1 and GSH or Trx. The p50 WT or C62S (0.05 μM) was incubated with the indicated concentrations of APE1/Ref-1, GSH and Trx/TrxR/NAPDH for 30 min, and then EMSA were performed with NF-κB probe. (B) The p50 (0.05 μM) was incubated with 0.5 μM APE1/Ref-1 WT or C/S with or without GSH for various times, and the redox state of p50 was visualized using F5M, as described in Materials and methods section. Fluorescence intensities of p50 were measured and plotted against time as fold change from the intensity of p50 incubated with GSH alone for 30 min. Plotted data are means ± SD from three independent experiments. (C) The p50 WT or C62S (0.05 μM) was incubated with 0.5 μM APE1/Ref-1 with or without GSH for 30 min, and then fluorescence assays were performed as in (B). (D) The p50 (0.05 μM) was incubated with the indicated concentrations of APE1/Ref-1 with or without Trx/TrxR/NAPDH for 30 min, and then fluorescence assays were performed as in (B).