Fig. 5.

Time course of Dot1a-Af9 epigenetic reprogramming of the αENaC R3 subregion in mIMCD3 cells treated with aldosterone. mIMCD3 cells were cultured in DMEM/F12 plus 10% charcoal-stripped serum for 16 h and then treated with vehicle or 1 μM aldo for the indicated time periods. The cells were then harvested and subject to ChIP/qPCR assays with the antibodies specific for Af9, Dot1a, the mineralocorticoid receptor (MR), or RNA polymerase II (RNA pol II). Specific primers were used to amplify the R3 subregion of αENaC. Data from the aldo-treated samples were normalized to the vehicle time control values, which did not significantly vary over the 3 h. The data are presented as the fold change of the indicated proteins relative to their value at time 0. Error bars indicate ± SE, n = 3. *P value ≤ 0.05 vs. corresponding value at time 0.