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. 2011 Sep 29;40(3):1251–1266. doi: 10.1093/nar/gkr791

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© The Author(s) 2011. 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 5. — Upf1 serine residue 1096 is necessary for the binding of the SMG-5:SMG-7 complex. (A) Schematic representation of Upf1 mutants. The NCR, cystein rich region, helicase domains and SQ-rich region are depicted by gray boxes. In Upf1-S1078A, S1078 is substituted for A. In Upf1-S1096, S1096 is replaced by A. In Upf1-S1116A, S1116 is substituted for A. In, Upf1-T28A, T28 is replaced by A. (B and C) HeLa TetOff cells were transfected with the indicated plasmids or an empty vector (‘vector’). The cell extracts were purified with streptavidin sephorose in the presence of RNaseA. Purified fractions were analyzed by western blotting with the antibodies indicated. (D) HeLa TetOff cells were transfected with the indicated siRNAs. The cell extracts were immunoprecipitated with anti-Upf1 (5C3) antibody in the presence of RNaseA. The immunoprecipitates (IP) were analyzed by western blotting with the antibodies shown on the left of the panels.