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. 2010 May 27;6(5):e1000920. doi: 10.1371/journal.ppat.1000920

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Braun et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Tg-AGO complexes were immunoprecipitated using Flag antibodies. The co-immunoprecipitated proteins were analysed using mass spectrometry (see Table 1 and Supplemental Table S1). Molecular weight markers are indicated on the left. (B) Transgenic parasites expressing ectopically HAFlag-TgAGO^FL were lysed and whole-cell-extracts prepared in the absence or presence (+RNase T1) of ribonuclease T1. HAFlag-TgAGO^FL was immunoprecipitated using anti-FLAG beads and flag peptide-eluted fractions were separated by SDS-polyacrylamide gel electrophoresis (4% to 12%), and visualized by silver staining. Molecular weight markers are indicated on the left. (C) Flag-immunopurified fractions from HAFlag-TgAGO^FL and HAFlag-TgAGO^DRGG RNase T1-treated and untreated samples were analyzed by Western blot using a home-made anti-Tg-KSRP antibody. (D) Sedimentation characteristics of Tg-KSRP compared to HAFlag-TgAGO^FL.