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. Author manuscript; available in PMC: 2014 Dec 30.

Published in final edited form as: Mol Carcinog. 2009 Feb;48(2):141–149. doi: 10.1002/mc.20462

In vivo interaction of c-Myc with the c-Myc site in AR gene and ET-1-induced expression of AR regulatory genes. (A) & (B) Chromatin immunoprecipitation (CHIP) assays showing in vivo interaction of c-Myc and c-myc binding region. Complexes of DNAs and transcription factors from LNCaP cells treated without and with ET-1 were subjected to immunoprecipitated with antibodies of c-Myc and control IgG. The purified DNAs in the supernatants (input) and imunnoprecipitates were amplified by PCR with primers specific to c-Myc (A) and control primers (B) (see Methods). Lane 1 is 1 kb DNA marker (Fermentas life science, MD). Lanes 2, 3 and 4 are the CHIP products from input, rabbit IgG control and c-Myc antibody with PBS treatment. Lanes 5, 6 and 7 are the CHIP products from input, rabbit IgG control and c-Myc antibody with 50 nM ET-1 treatment for 18 hrs. The data represent one experiment performed on three separate occasions with similar results. (C) Cell lysates of LNCaP cells treated with 50 nM ET-1 or pre-exposure to NEP and ET-1 antagonists BQ123 or BQ 788 were subjected to Western analysis and probed with antibodies of PSA and actin.

In vivo interaction of c-Myc with the c-Myc site in AR gene and ET-1-induced expression of AR regulatory genes. (A) & (B) Chromatin immunoprecipitation (CHIP) assays showing in vivo interaction of c-Myc and c-myc binding region. Complexes of DNAs and transcription factors from LNCaP cells treated without and with ET-1 were subjected to immunoprecipitated with antibodies of c-Myc and control IgG. The purified DNAs in the supernatants (input) and imunnoprecipitates were amplified by PCR with primers specific to c-Myc (A) and control primers (B) (see Methods). Lane 1 is 1 kb DNA marker (Fermentas life science, MD). Lanes 2, 3 and 4 are the CHIP products from input, rabbit IgG control and c-Myc antibody with PBS treatment. Lanes 5, 6 and 7 are the CHIP products from input, rabbit IgG control and c-Myc antibody with 50 nM ET-1 treatment for 18 hrs. The data represent one experiment performed on three separate occasions with similar results. (C) Cell lysates of LNCaP cells treated with 50 nM ET-1 or pre-exposure to NEP and ET-1 antagonists BQ123 or BQ 788 were subjected to Western analysis and probed with antibodies of PSA and actin.