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Figure 2 | Epigenetics & Chromatin

Figure 2

From: ERG-associated protein with SET domain (ESET)-Oct4 interaction regulates pluripotency and represses the trophectoderm lineage

Figure 2

ERG-associated protein with SET domain (ESET)-mediated histone 3 lysine 9 trimethylation (H3K9me3) represses Cdx2 in embryonic stem (ES) cells. (a) Chromatin immunoprecipitation (ChIP) primers C1 to C10 used to detect enrichment of H3K9me3 on Cdx2 promoter (left). Primers O1 of Oct4 promoter served as a negative control (right). The numbers below the bars indicate distance from transcription start site (TSS) in base pairs (bp). (b) Quantitative polymerase chain reaction (Q-PCR) analysis of the enrichment of H3K9me3 at different positions along Cdx2 promoter region relative to the Oct4 promoter region after normalising against H3 ChIP and IgG controls. Primers C4 and C6 were not suitable for Q-PCR analysis. Error bars, standard deviation (SD) of three technical replicates. (c) Carrier ChIP semiquantitative PCR analysis of H3K9me3 at the Cdx2 promoter and major satellite regions of fluorescence-activated cell sorting (FACS)-sorted Eset knockdown ES cells relative to control cells after normalising against their respective input and IgG controls. Primers C6 and C7 were not suitable for carrier ChIP analysis. Error bars, SD of three independent experiments. (d) Q-PCR analysis of the levels of H3K9me3 at region C1 and C2 of the Cdx2 promoter in Eset knockdown ES cells relative to control cells after normalising against their respective input and IgG controls. Error bars, SD of three independent experiments. (e) Q-PCR analysis of the enrichment of ESET on different positions along the Cdx2 promoter relative to the Oct4 promoter after normalising against their respective input and IgG controls. Error bars, SD of three technical replicates.

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