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. 2014 Sep 15;8(6):1845–1856. doi: 10.1016/j.celrep.2014.08.020

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© 2014 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).

PMC Copyright notice

DHX34 Promotes eRF3 Release from the SURF Complex

(A) FLAG-UPF1 was transiently coexpressed with T7-DHX34 in HEK293T cells. Inputs (0.5%) and anti-FLAG IPs (20%) were analyzed with the indicated antibodies.

(B) HEK293T cells were transiently transfected with FLAG-UPF1 and with an shRNA targeting DHX34 or with an empty vector control. Inputs (0.5%) and anti-FLAG (20%) IPs were analyzed by western blotting for the indicated proteins.

(C and D) Cells were transfected with an shRNA targeting DHX34 or with an empty vector control. After 4 days cells were cotransfected with FLAG-eRF1 (C) or FLAG-eRF3 (D) and shRNA-resistant (^R) T7-DHX34 (wild-type or the ATPase-deficient mutants K191S and D279A). Anti-FLAG immunoprecipitations were performed 2 days later and analyzed for the presence of DHX34. Input (0.5%) and anti-FLAG IPs (20%) were analyzed by western blotting with the indicated antibodies.