Thinned apple polyphenols (TAP) extract was evaluated for its ability to modulate the antioxidant response mediated by NRF2 activation using NRF2/ARE Responsive Luciferase Reporter HEK293 stable cell line (Signosis, Santa Clara, CA, USA) as previously described [46 (link)]. Briefly, HEK293 cells (10,000 cells/well) were treated with the extract (concentrations between 1 and 250 µg/mL). CDDO-Me 75 (bardoxolone methyl) 75 nM was used as a positive control [47 (link)]. After adding ONE-Glo™ Luciferase Assay Substrate (purchased from Promega Corporation, Madison, WI, USA) (100 µL/well), luciferase measurement was performed with a luminometer (Wallac Victor2 1420, Perkin-Elmer™ Life Science, Monza, Italy). Experiments were carried out with biological and technical replicates; values are reported as mean ± SD compared to untreated control cells. One-way ANOVA with Bonferroni’s multiple comparisons test (p < 0.05 was considered significant) was used for the statistical analysis.
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