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. 2012 May 30;40(16):7676–7689. doi: 10.1093/nar/gks509

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© The Author(s) 2012. 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/3.0), 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. — HP1γ knockdown increases expression of TNF and abolishes H3.3S31 phosphorylation. (A) HP1γ, TNF and IKKα mRNA levels in RAW264.7 cells after HP1γ knockdown using three shRNAs against HP1γ (sh-1, -2 and -3) or firefly luciferase (sh-c) after 1 h LPS treatment. Results are expressed relative to GAPDH expression. Error bars represent standard deviation (SD) from three independent experiments. (B) Quantification of total HP1γ and β-actin protein levels by western blot with total cell extract, after HP1γ knockdown (sh-1) in RAW264.7 cells. (C,E) ChIP performed, after HP1γ knockdown in RAW264.7 cells treated with LPS for 1 h, with (C) anti-RNAPII S2p and (E) anti-H3.3S31p antibodies. Horizontal axis indicates primers used for the real-time PCR. Data are normalized versus input or (E) total histone H3 and then versus an average of background control regions. (D) qPCR quantification of MNase-treated chromatin. (C–E) Data are representative of at least three independent experiments. Error bars represent SD from three independent qPCR replicates.