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. 2009 Nov 11;38(2):382–390. doi: 10.1093/nar/gkp991

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© The Author(s) 2009. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Parafibromin recruits SUV39H1 to promote H3 K9 methylation around cyclin D1 gene. (A) Cyclin D1 locus showing regions subjected to ChIP analysis. (B, C) HeLa cells were treated with siRNA targeting HRPT2 or with an untargeted control siRNA for 24 h. Cells were then analyzed by ChIP using control antibody (rabbit IgG) and antibodies against indicated proteins or H3 modifications as described in ‘Materials and methods’ section. Immunoprecipitated DNA was analyzed in duplicates by quantitative RT–PCR to measure the relative recruitment of parafibromin, SUV39H1, or G9a (B) or the relative levels of H3K4 or H3K9 methylation along the cyclin D1 locus (C) ChIP with control IgG was subtracted from all ChIP values (IP/Input, the percentage of ChIP). Data represent the percentage of ChIP signal normalized to the value of promoter region of the control siRNA sample to compare the relative enrichment of signals along the region. (The ChIP value obtained by amplification of promoter region of the control siRNA sample was arbitrarily set to 1 in each panel.) Data represent the average of two or three independent experiments. Error bars indicate the standard deviation between experiments.