Fig. 2

MMP13 gene is an ETV4 target gene in mammary epithelial TAC and MMT cells. Effect of ETS/AP1 binding site mutations in the MMP13 promoter regulation. a Schematic representation of the mouse MMP13 promoter fragments (pMMP13–1800 to pMMP13–91) and AP-1 and/or ETV4 mutant versions cloned into a pG3bLuc reporter vector (pG3b). Position of the conserved ETS binding sites (EBS) is represented by . AP-1 binding site is represented by . : Transcription start site. : Mutation of the ETS site. b and c Histograms representing the relative luciferase activity measured for each promoter construct cotransfected into the TAC cell line with pTracer vector (−) or pTracer-ETV4 expression vector (ETV4) and/or AP-1 expression vector (AP1). Experiments were conducted three times in triplicate. Error bars indicate SD. d ChIP experiment. PCR detection of the MMP13 promoter region after ETV4 immunoprecipitation in MMT (left panel) and TAC (right panel). Primers allowing the amplification of the proximal MMP13 promoter region containing EBS are schematized in the lower panel. Cyclin D2 was used as a positive control [8]. Immunoprecipitation with a nonrelevant antibody (IgG) was used as a negative control.