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. 2005 Jul 26;33(13):4191–4201. doi: 10.1093/nar/gki734

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© The Author 2005. Published by Oxford University Press. All rights reserved

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(A) Genomic sequence of the human HOXA10 regulatory region. The GRE(HOXA10) sequence used for EMSA is indicated by a double underline. The primer sets used for the ChIP assays are also shown. (B) Binding of GZF1 to the HOXA10 regulatory region as assessed by ChIP. Crosslinked chromatin fragments from HEK293T cells expressing GZF1 or GFP-GZF1 were immunoprecipitated with anti-GZF1 antibody or anti-GFP antibody. DNA from the resulting immunoprecipitates was subjected to PCR amplification using the primers shown in (A). As a positive control (input), 10 ng of genomic DNA was amplified in parallel. (C) Binding of GZF1 to GRE(HOXA10) as assessed by EMSA. EMSA was performed using a ³²P-labeled GRE(HOXA10) probe and GST-ΔBTB-GZF1 protein (0.1 and 0.3 μg). The GRE(HOXA10) probe and GST-ΔBTB-GZF1 protein showed a single complex band, which was identical to that shifted with the GRE probe.