Figure 8

Overexpression of active NRF2 partially prevents NOX4 deletion-associated pro-apoptotic phenotype.

(A) Real time PCR analysis of NRF2 mRNA and (B) Western blot analysis of NRF2 protein and β-actin performed on WT MEF cells transfected or not with active NRF2. Representative images are shown. Results are expressed as ratio of relative quantity over the mean value obtained in WT control cells ± SEM, (n = 6), ns p > 0.05, * or ^#p < 0.05. (C) PCR analysis of GSTα2, NQO-1 and GCLC (E) mRNA expression performed on WT and NOX4 KO MEF cells transfected or not (vector) with active NRF2 (n = 6). Results are expressed as ratio of relative quantity over the mean value obtained in WT control cells ± SEM, (n = 6), ns p > 0.05, *p < 0.05, ^##p < 0.01, ^###p < 0.001. (D) Western blot analysis of NRF2, Bcl-2 and β-actin performed on WT and NOX4 KO MEF cells transfected or not with active NRF2. Representative images are shown and quantification (E) of Bcl-2 expression (n = 3). Results are expressed as ratio of relative quantity over the mean value obtained in WT control cells ± SEM, (n = 3), ns p > 0.05, *p < 0.05.(F) Measurement of total glutathione in WT and NOX4 MEF cells transfected or not with active NRF2. Results are expressed as ratio of relative quantity over the mean value obtained in WT control cells ± SEM, (n = 3), ns p > 0.05, *p < 0.05. (G) Annexin V/PI assay on WT and NOX4 KO MEF cells transfected or not with active NRF2. Results are expressed as percentage of late apoptotic cells in FACS analysis. (n = 3), ns p > 0.05, *p < 0.05.