ChIP assays to demonstrate that ETS-1, CREB, AP-2α and SP-1 bind to PP-2α promoter.

<p>ChIP assays were conducted as we recently described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007019#pone.0007019-Qin1" target="_blank">[34]</a>. The oligos used for ChIP assays were described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007019#s2" target="_blank">Materials and Methods</a>. A. ChIP assays to show that ETS-1 and CREB bind to PP-2Aα promoter. Lane 1, DNA marker; lane 2, PCR product derived from 1/50 DNA template immunoprecipitated by anti-CREB antibody; Lane 3, PCR product derived from DNA template immunoprecipitated by normal IgG; lane 4, PCR product derived from 1/50 DNA template immunoprecipitated by anti-ETS-1 antibody; Lane 5, PCR product derived from DNA template immunoprecipitated by normal IgG; lane 6, PCR product derived from direct input DNA template without immunoprecipitation. A band of 189 bp containing both CREB and ETS-1 binding sites in the PP2A-Aα promoter gene was amplified. ChIP assay confirms that CREB and ETS-1 bind to PP2A-Aα gene. B. ChIP assays to show that AP-2α and SP-1 bind to PP-2Aα promoter. Lane 1, DNA marker; lane 2, PCR product derived from 1/50 DNA template immunoprecipitated by anti-SP-1 antibody; Lane 3, PCR product derived from DNA template immunoprecipitated by normal IgG; lane 4, PCR product derived from 1/50 DNA template immunoprecipitated by anti-AP-2α antibody; Lane 5, PCR product derived from DNA template immunoprecipitated by normal IgG; lane 6, PCR product derived from direct input DNA template without immunoprecipitation. A band of 168 bp containing both AP-2α and SP-1 binding sites in the PP2A-Aα promoter gene was amplified. ChIP assays confirm that AP-2α and SP-1 bind to PP2A-Aα gene.