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2-ME activates PKR in a cell-free system. Cytoplasmic extracts prepared from untreated MG63 cells were incubated in kinase assay mixture with vehicle (Veh) or 2, 10, and 50 µM 2-ME and assayed for PKR activity in vitro. The kinase assay mixture was analyzed by Western blot hybridization using anit-phospho-PKR (pT446; Epitomics) and anti-eIF-2α (Cell Signaling Technology).
