Figure 7

Analysis of Sp1 binding to the ACVR1 72 - bp promoter region. The Sp1 transcription factor binds the ACVR1 basal promoter as assessed by EMSA experiments with HeLa nuclear extracts and a double-stranded probe spanning the GC-boxes contained in the fragment (position -38 to -9). Lane 1, Free probe; lane 2, probe incubated with HeLa nuclear extracts (NE); lanes 3 to 5, DNA-protein complexes (arrows) competed with the cold wild-type probe (50-, 100-, or 200-fold molar excess, respectively); lanes 4 to 8, DNA-protein complexes competed with the cold mutated probe (50-, 100-, or 200-fold molar excess, respectively); lanes 9 and 10, DNA-proteins complexes incubated with probes containing canonical consensus sites for Sp1 and Egr-family factors respectively; lanes 11 to 13, super-shift analyses of the DNA-protein complexes with three different specific antibodies against Sp1. *, retarded complexes recognized by anti-Sp1 antibodies.