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Figure 5 | Epigenetics & Chromatin

Figure 5

From: PAT-ChIP coupled with laser microdissection allows the study of chromatin in selected cell populations from paraffin-embedded patient samples

Figure 5

Application of PAT-ChIP to human LMD samples. Six tissue slides (6 μm thick) were prepared from both human FFPE lung adenocarcinoma (AC) and FFPE lung squamous carcinoma (SC) tissues and subjected to LMD to isolate normal and tumor cells. Chromatin was then immunoprecipitated with anti-H3K4me3 and anti-H3K27me3 antibodies, and the resulting purified DNA was analyzed by real-time qPCR for enrichment at specific loci. (A) Representative H&E staining of the two human FFPE lung samples. (B) Evaluation of chromatin fragmentation by electrophoretic separation on 1.3% agarose gel and SYBR® Gold staining of DNA purified only from H3K4me3 unbound fractions. (C) Amplification of transcriptionally active (Vcl and Gapdh) and inactive (Hapln1 and Col2a1) promoter regions by real-time qPCR (each sample amplified in triplicate). Enrichments of the promoter sequences associated with the indicated genes for H3K4me3 (n.r. Ab, non-related antibody) are expressed as the ratio between bound and input (percentage).

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Epigenetics & Chromatin

ISSN: 1756-8935