FIGURE 8.

HDACi-mediated in vivo regulation of renal NPRA expression and signaling involving HDACs, acetylated histones, Sp1, and p300 in Npr1 gene-targeted mice. A, effect of HDACi on renal HAT activity and global acetylation levels of H3-K9ac and H4-K12ac. B, HDACi-mediated recruitment of acetylated histones, HDAC1 and -2, Sp1, and p300 at the Npr1 promoter (−120 to +73) in treated mice kidneys. Pooled samples from 6 mice in each group were used for ChIP and purified DNA was amplified by conventional PCR. C, Npr1 mRNA levels in TSA- and MGCD0103-treated mice kidneys as analyzed by real-time RT-PCR with β-actin as an internal control. D, Western blot and densitometry analyses of NPRA protein expression in TSA- and MGCD0103-treated Npr1 1-copy, 2-copy, and 4-copy mice kidneys. β-Actin was used as loading control. E, cGMP levels in kidney tissues of HDACi-treated mice as quantitated by ELISA. Data shown represent mean ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (vehicle-treated versus drug-treated same group). Ab, antibody. In each group, six mice (n = 6) were used.