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. 2009 Nov 11;38(2):360–369. doi: 10.1093/nar/gkp977

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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 5. — MePCE not bound to LARP7 are responsible for all the 7SK capping activity in the nucleus and preferentially associated with the 7SK gene promoter. (A) HeLa NEs were subjected to immunodepletion with the indicated antibodies and then analyzed by WB for the presence of the indicated proteins (right panel). The depleted NEs were then tested in 7SK capping reactions, with the RNA products analyzed by autoradiography (top left) and EB staining (bottom left). (B) The levels of input chromatin derived from the HeLa-based W10 cells stably expressing F-MePCE were carefully adjusted (in 2-fold increments) and then PCR amplified with primer sets corresponding to the indicated genes (top panel). Once the input chromatin corresponding to the five genes were normalized to approximately the same level, ChIP with anti-Flag mAb was performed. After DNA purification, PCR reactions containing α-[³²P]dCTP were carried out and the products analyzed by gel electrophoresis and autoradiography (bottom panel).