Fig. 4.

Examples of predicted TFBSs and the impact of transcription factors on BMP4 target gene expression. a The predicted binding sites of transcription factors MBD2, HIF1A and CBFB are depicted at the promoters of NOG, SMAD7 and ID1 genes, respectively. In addition, known BMP-response elements (BRE) located near the binding sites are illustrated. b The TFs were silenced and the cells were treated with BMP4 or vehicle control followed by measurement of target gene expression by qRT-PCR. Examples of relative expression levels of SKIL after HIF1A silencing in MDA-MB-231 cells (top panel) and NOG expression after MBD2 silencing in T-47D cells (bottom panel) are shown. c Graphical summary of the TF silencing experiments. The order of the genes is identical to that in Fig. 2. Blue color (decreased target gene expression) denotes TFs that were essential for target gene expression and red color those whose silencing led to enhanced target gene expression. Not applicable indicates cases where BMP4 did not alter the baseline gene expression. Data on the DLL1 gene, which is downregulated in T-47D upon BMP4 treatment, are highlighted with a bold line