Figure 1. Runx1 binds to Rspo3 promoter.

(A) ChIP assays were performed on LM3 cells using specific ChIP-grade Runx1 antibody or control IgG antibody. Specific primers were designed for targeting Runx1 high affinity binding site in the Rspo3's promoter region. Bar graph shows mean and standard deviation (SD) of three independent experiments each of them performed by triplicate. Primers for Gapdh promoter region were used as negative control with no amplification product. (B–C) Gel shift assays were performed on LM3 (B) or SCp2 (C) nuclear extracts using an oligoprobe containing Runx1 consensus sequence included in the Rspo3 promoter region (−490bp) (lane ³²P and lane Ab). This band showed lower intensity when cold oligonucleotides were included in the reaction (lane C). Asterisks in the Figure show unspecific binding. ³²P: phospho-labeled oligoprobe, Ab: anti-Runx1 antibody and C unlabeled oligoprobe.