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. 2017 May 17;8(37):60809–60825. doi: 10.18632/oncotarget.17955

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Copyright: © 2017 Zimmermann et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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A-C. Identification of over-represented transcription factor binding sites (TFBS) in sets of differentially expressed genes in myocardium by oPOSSUM. A. In the infarct core zone members of the ETS transcription factor family (blue diamonds), key regulators of the hypertrophic transcriptional program (orange diamonds), signaling transducer and activator of transcription 3 (STAT3, green diamond) and kruppel-like factor 4 (Klf4, red diamond) binding sites were significantly enriched within the promoter regions. B. and C. In the border and remote zone AT-Rich Interaction Domain 3 (ARID3A, green diamond) and Nkx2-5 (red diamond) binding sites were significantly enriched within the promoter regions. D. Rank test of selected transcription factors based on the Fisher exact test score for different myocardial regions (IZ…infarct core zone, BZ … border zone, RZ … remote zone). A rank% value of 100 indicates that the TF mentioned are most probably functionally relevant. A rank% value of < 70 indicates that these TF may not be involved in gene regulation. Klf4 shows high values in the IZ and low values in the BZ and RZ. E. Five different down-stream targets of Klf4 show a significant up- (DERL3) and downregulation (ACTA1, ACTC1, DSTN, GATA4) of mRNA-levels in the infarct core-, border- and remote zone as compared to the control zone, measured by RT-PCR. Gene expression levels were calculated based on the mean value from six samples using the comparative Ct method and normalized to the housekeeping gene beta actin (ACTB). Data are presented as mean + SD.