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. 2017 Sep 11;9(9):1001. doi: 10.3390/nu9091001

Figure 1.

Figure 1

Effects of hydroxy fatty acids on Nrf2 translocation and expression of genes regulated by the Keap1-Nrf2 pathway in HUVECs. Nrf2 translocation was detected by immunostaining (green), and the nuclei were stained with PI (red). Cells were treated for 3 h with: 0.5% MeOH (vehicle) (A); 10 μM tBHQ (B); 50 μM AA (C); 50 μM EPA (D); 50 μM 5-HETE (E); and 50 μM 5-HEPE (F). The intensity of the green fluorescence signal in the nucleus was quantified using Image J (G). Data are expressed as the mean ± standard deviation (SD) (n = 10). Significant differences among the groups are indicated with different letters (one-way ANOVA followed by Tukey’s test, p < 0.05). Gene expression levels of: HMOX1 (H); and SLC7A11 (I) in HUVECs were analyzed and normalized to 18S gene expression levels (as an internal control) after incubation of each sample for 6 h. Data are expressed as mean ± SD (n = 4). Significant differences among the groups are indicated with different letters (one-way ANOVA followed by Tukey’s test, p < 0.05). Abbreviations: Nrf2, NF-E2 related factor 2; Keap1, Kelch-like ECH-associated protein 1; HUVEC, human umbilical vein endothelial cell; PI, propidium iodide; MeOH, methanol; tBHQ, tert-butylhydroquinone; AA, arachidonic acid; EPA, eicosapentaenoic acid; 5-HETE, 5-hydroxyeicosatetraenoic acid; 5-HEPE, 5-hydroxyeicosapentaenoic acid; ANOVA, analysis of variance; and HMOX1, heme oxygenase 1.