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. 2013 Nov 1;19(13):1507–1521. doi: 10.1089/ars.2012.4713

FIG. 2.

FIG. 2.

SirT1 is recruited to promoter regions of ROS detoxification genes upon serum starvation. (A–E) Chromatin immunoprecipitation assays of promoter regions corresponding to the indicated ROS detoxification genes. Cross-linked chromatin Promoter occupancy was analyzed by qPCR using specific primers. β-actin CDS was used to control for non specific enrichment. (A) SirT1 binding sites. (B) H4K16 acetylation levels at SirT1 sites. (C) Pol II recruitment. The position of the analyzed fragments is indicated, using as reference (+1), the position of the ATG. (D, E) Analysis of prx3 and ucp-2 genes. Left panels, analysis of Pol II at the transcriptional start region and along the gene. Central panel, analysis of “elongating” Pol II levels (Pol II phosphorylated in the Ser2 positions of the CTD repeats of the Pol II largest subunit) at the transcriptional start region and downstream of the ATG. Right panels, SirT1 recruitment along the genes, upstream and downstream of the ATG. The position of the analyzed fragments is indicated, using as reference (+1), the position of the ATG. (F) mRNA and (G) protein levels of SirT1 target genes in confluent BAEC cells in 10% fetal bovine serum (FBS) or serum deprived (0.5% FBS) o/n were analyzed by qRT-PCR and western blot, respectively. Data are from ≥3 independent experiments. Data are means+SD (*) p≤0.05 versus control. The blots presented correspond to a representative experiment.