PMC

(A) Sequencing of genomic DNA of wild type (WT; n=3) and p67^phox−/− revealed a 5 bp deletion (GAGAA) in the genomic sequence of p67^phox−/−. (B) Respiratory burst of macrophages isolated from p67^phox−/− (n=6) and WT (n=4) rats in response to phorbol 12-myristate 13-acetate (PMA), a PKC activator, administered at time zero.

Salt-sensitive hypertension and renal oxidative stress are significantly attenuated in the p67^phox−/− rats compared to wild type (WT) littermates. (A) Mean arterial pressure (MAP) of 5–6 week-old rats on 0.4% and 8% salt diet (n=6–9 rats/strain). † p<0.001 significant difference between strains at that time point; (B) outer medullary NAD(P)H oxidase on D14-HS (n=6–9 rats/strain); (C) outer medullary superoxide on D14-HS (n=6–9 rats/strain); (D) renal interstitial H₂O₂ (n=5–8 rats/strain). * p<0.05 significant difference from WT.

Greater p67^phox expression is associated with greater NAD(P)H oxidase activity and salt-sensitivity in the SS rat. (A) Western blot densitometric analyses of p67^phox (β-actin normalized) of rats on both 0.4% LS and day 7 of HS diet with the associated gel shown (n=5 rats/strain/time point). * p<0.05 significant difference from SS. (B) NAD(P)H oxidase activity of rats on both LS and D7-HS (n=11–12 rats/strain/time point). * p<0.01 significant difference from LS within strain; † p<0.01 significant difference from SS. (C) Mean arterial pressure (MAP) of 5–6 week-old rats on both 0.4% and 8% salt diet (n=8–10 rats/strain). † p<0.001 significant difference between strains at that time point.

Genetic variances in the promoter region of p67^phox affect promoter activity and gene expression. (A) Quantitative real-time PCR of p67^phox of rats (N=6 per strain at each time point) on both 0.4% low salt (LS), and day 7 of 8% high salt (D7-HS). * p< 0.05 significant difference from SS. (B) Schematic of constructed SS and SS.13^BN26 p67^phox promoters. The SS p67^phox promoter not only contains 204bp deletions, but also has 4 SNPs downstream of the deletion. (C) Luciferase activity of both SS and SS.13^BN26 p67^phox promoters (n=8–9 per strain). * p<0.05 significant difference from SS.

Renal injury is significantly reduced in p67^phox−/− rats compared to wild type (WT) littermates. (A) Proteinuria and (B) microalbuminuria of p67^phox−/− and WT rats on LS, D7-HS, and D14-HS diet (n=6 rats/strain). † p<0.01 significant difference from WT at the same time point; (C), (D), (E) Histological staining and quantitative analyses of renal injury markers in p67^phox−/− and WT littermates on D14-HS diet. (C) Trichrome staining for the quantification of protein casts (n=7); (D) α-SMA immunostaining for fibrosis (n=6), and (E) ED-1 immunostaining (n=5); (F) Cortical glomerular injury of superficial glomeruli (n=7). *p<0.05 significant difference from WT.