Figure 5: Association of LSF with p21CIP1 promoter sequences in vitro and in vivo. (a) Putative CNRG-N6-CNRG motifs in the p21CIP1 promoter region. (b) EMSA assay performed with putative LSF-binding and mutant DNA probes in the presence (+) or absence (-) of in vitro translated LSF and competitor oligonucleotides. (c) A luciferase reporter construct containing the p21CIP1 promoter was transfected with or without LSF siRNA in parental SK-Mel28 cells or in cell clones that stably overexpress the DsRed-LSF fusion protein or empty vector. (d) ChIP assay indicated that LSF is associated with the two promoter regions of p21CIP1 as shown by the enrichment detected by qPCR at these sites following precipitation with an anti-DsRed antibody compared with IgG-precipitated chromatin. All charted data represent mean ± SD of three independent experiments. **P<0.01. Gray bars in gene map represent position of each probe.