FIGURE 7.

Nox4 activates c-Jun in NRCs but does not affect GATA-4 mRNA expression levels. A, representative Western blot analyses of Nox4 protein levels in whole cell lysates or phospho- and total c-Jun protein levels in nuclear extracts of NRCs transduced with either AdβGal or AdNox4 for 24 h. Actin is included as a loading control. B, densitometric analysis of the proportion of c-Jun phosphorylation, derived from three separate Western blot analyses. C, ELISA-based analyses of binding activity of c-Jun in nuclear extracts of NRCs transduced with AdβGal-transduced (open bar) or AdNox4-transduced (black bar) as in A. D, luciferase activity resulting from NRCs transduced for 24 h with either AdβGal or AdNox4 as indicated and subsequently transfected with an AP-1-luciferase reporter construct (black bars) or minimal promoter-containing control plasmid (open bars) and cultured for a further 24 h. Luciferase activity was normalized to that from the co-transfected SV40 Renilla luciferase control vector and is expressed as RLU. E, Q-PCR analysis of GATA-4 mRNA levels in NRCs transduced with AdβGal or AdNox4 for 24 h. F, representative Western blot analyses of GATA-4 protein levels in whole-cell lysates of NRCs transduced with either AdβGal or AdNox4 for 24 h. Actin is included as a loading control. All data are presented as mean ± S.E. *, p < 0.05; n.s., not significant.