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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Cancer Res. 2008 Nov 1;68(21):8770–8778. doi: 10.1158/0008-5472.CAN-08-1912

Figure 5.

Figure 5

Recruitment of HDAC-1 to NKX3.1 binding site of VEGF-C promoter. A, ChIP: cross-linked chromatin-protein complexes were isolated from regular LNCaP cells or LNCaP cells transfected with different doses of siRNA against NKX3.1. Immunoprecipitation of chromatin fragments was carried out with the HDAC-1 as well as rabbit IgG (control) antibody. After reverse cross-linking, DNA fragments were isolated and the NKX3.1 binding regions (−997, −1614, and −2359 bp) on the VEGF-C promoter were amplified by PCR with the VEGF-C promoter–specific primers. Only −997 bp was positive. Re-ChIP: B, re-ChIP with anti-HDAC1 antibody as well as rabbit IgG (control) antibody was performed with the cross-linked protein-DNA complex eluted from the first step ChIP with NKX3.1 antibody as described in Fig. 3A. C, re-ChIP with anti-NKX3.1 antibody was performed with the cross-linked protein-DNA complex eluted from the first step ChIP with anti-HDAC1 antibody. Re-ChIP with rabbit IgG was used as control.