Chromatin changes at the AR gene in response to hypertonic stress.
- Publication date
- Publisher
Abstract
<p>(A-C) NIH-3T3 cells were subjected to hypertonic stress, and ChIP was conducted with antibodies against (A) histone H3, (B) histone H2B, and (C) histone H3. DNA was analyzed by real-time PCR using AR<sub>INTER</sub> (a), AR<sub>ORE</sub> (b), AR<sub>PP</sub> (c), and AR<sub>EX2</sub> (d) primer pair respectively as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0008435#pone-0008435-g001" target="_blank">Figure 1</a>. *, p<0.01 and **, p<0.001 by one-way ANOVA. (C) Cells were either left untreated (isotonic), or treated with hypertonic medium for 30 min followed by incubation in isotonic medium for 30 min, 6 and 16 h respectively (isotonic recovery). ChIP was conducted with antibody against histone H3. DNA was analyzed by real-time PCR using AR<sub>ORE</sub> primer pair. The results are presented as the ratio of immunoprecipitated DNA to total input DNA and normalized to the value of cells maintained under isotonicity. *, p<0.001 by one-way ANOVA. (D) Cells were subjected to hypertonic stress, and ChIP was conducted with antibodies against OREBP. DNA was analyzed by real-time PCR using AR<sub>ORE</sub>. The results were normalized to the ratio of immunoprecipitated DNA to total input DNA at time = 0. (E and F) Cells were treated with hypertonicity for 0 and 8 hr, respectively. ChIP was conducted with antibodies against OREBP (E) or histone H3 (F). DNA was analyzed by real-time PCR using a primer pair specific for the TonEA of the SMIT gene. The results were normalized to the ratio of immunoprecipitated DNA to total input DNA at time = 0. *, p<0.05 by Student's t-test. For A-G, data are the mean ± SEM (n = 3).</p
