ChIP-qPCR Analysis of Histone Modifications

ChIP analysis was performed as previously reported [27 (link)] by using 5 μg of the following antibodies: rabbit anti-H3K4me3 (07473), rabbit anti-H3K27me3 (07449), rabbit anti-H3AcK9 (07352), or rabbit IgG as a negative control (12307) (all from Millipore, MA, USA). 5 ng of immunoprecipitated DNA and the relative control were used as templates for RT-qPCR analysis, performed in duplicate. Primers utilized to amplify the HNF1/6 consensus on HNF4α promoter are as follows: forward 5′-TCCGAAAGACCCAAGTGTGG-3′ and reverse 5′-GCCAATCACGTCCCAGATCA-3′.

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Cozzolino A.M., Noce V., Battistelli C., Marchetti A., Grassi G., Cicchini C., Tripodi M, & Amicone L. (2016). Modulating the Substrate Stiffness to Manipulate Differentiation of Resident Liver Stem Cells and to Improve the Differentiation State of Hepatocytes. Stem Cells International, 2016, 5481493.

Publication 2016

rabbit anti-H3K4me3 (07473 rabbit anti-H3K27me3 (07449 rabbit IgG

Antibodies Chip H3k4me3 Primers Rabbit

Corresponding Organization : Istituti di Ricovero e Cura a Carattere Scientifico

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Variable analysis

independent variables

Antibodies used for ChIP analysis: rabbit anti-H3K4me3, rabbit anti-H3K27me3, rabbit anti-H3AcK9, or rabbit IgG as a negative control

dependent variables

Enrichment of histone modifications (H3K4me3, H3K27me3, H3AcK9) at the HNF1/6 consensus on the HNF4α promoter, as measured by RT-qPCR

control variables

Amount of immunoprecipitated DNA used as template for RT-qPCR (5 ng)
Primers used to amplify the HNF1/6 consensus on the HNF4α promoter

controls

Rabbit anti-H3K4me3, rabbit anti-H3K27me3, rabbit anti-H3AcK9 antibodies
Rabbit IgG

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