1. ZEB1‐dependent epigenetic modifications ASchemes for the genomic loci of the miRNA and E‐cadherin genes, showing regions of the CpG islands (yellow), of the qRT–PCR amplicon for chromatin immunoprecipitation (ChIP) analysis (blue) and of the bisulfite sequencing (red).B, CHistone marks were analyzed using ChIP coupled to qRT–PCR for Panc1 control versus shZEB, MDA‐MB‐231 control versus shZEB (B), and BxPC3 control versus gemcitabine resistant (gr) (C).
ZEB1‐dependent epigenetic modifications ASchemes for the genomic loci of the miRNA and E‐cadherin genes, showing regions of the CpG islands (yellow), of the qRT–PCR amplicon for chromatin immunoprecipitation (ChIP) analysis (blue) and of the bisulfite sequencing (red).B, CHistone marks were analyzed using ChIP coupled to qRT–PCR for Panc1 control versus shZEB, MDA‐MB‐231 control versus shZEB (B), and BxPC3 control versus gemcitabine resistant (gr) (C). In MDA‐MB‐231 and Panc1, the active histone marks H3K4me3, H3ac, H4ac, and H3K9ac were enriched. Vice versa, in the drug‐resistant clones of BxPC3, the active marks were reduced in the CpG islands. The repressive histone mark H3K27me3 was not detectable in the miR‐200 loci, but in the loci of miR‐203 and E‐cadherin in Panc1 and MDA‐MB‐231. DNA methylation status was determined by bisulfite sequencing. Depletion of ZEB1 in MDA‐MB‐231 resulted in almost complete demethylation, whereas the selection of drug‐resistant, ZEB1‐expressing clones in BxPC3 induced complete methylation. n = 2 (Panc1) or 3 (MDA‐MB‐231 and BxPC3), mean ± SEM; unpaired Student's t‐test.
Simone Meidhof et al. EMBO Mol Med. 2015;7:
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