Figure 2

A functional GATA-1 element in the proximal ALCAM promoter. A) Schematic diagram of reporter constructs consisting of truncated ALCAM promoter fragments driving expression of pGL3. Histogram shows the relative luciferase activity for each construct in K562 and MEG-01 cells. B) Reporter constructs showing the presence of wild-type (filled circle) or mutant (open circle) GATA-1 cis element at -850 in the human ALCAM promoter. Histogram shows the relative luciferase activity for each construct in K562 and MEG-01 cells. C) Electrophoretic mobility shift assay showing protein-DNA complexes in experiments performed with the -850 GATA probe and no nuclear extracts (lane 1), or with K562 nuclear extracts alone (lane 2), and in the presence of molar excess of unlabeled wild-type probe (lanes 3-5), anti-GATA1 antibody (lane 6) and unlabeled mutant -850 GATA1 probe (lanes 7-9). D) Ethidium bromide stained gel of ChIP assay PCR products showing a 151 bp product for input DNA (lane 2), and products from chromatin immunoprecipitations performed with anti-GATA1 antibody (lane 3), IgG (lane 4) and without chromatin (lane 5). Lane 1 contains DNA size marker.