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Btk constructs were expressed in HEK293 cells using pCS2-gateway plasmid containing an N-terminal 6xMyc tag, while repebodies were cloned into pcDNA3.1 vector and contained a C-terminal Flag tag. Transient transfections with respective plasmids were performed using Polyfect transfection reagent (Qiagen). 48 h after transfection, cells were harvested, lysed and samples further processed for immunoblotting.
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