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R., & amp
10 g GST or GST-tagged proteins were bound to 20 l Glutathione Sepharose 4B beads (GE Healthcare) and incubated with 5 l purified HA-Atg8a or 5 l PURExpress reaction mixture for 2 h at 4C in binding buffer (50 mM Hepes, pH 7.5, 150 mM NaCl, 2 mM MgCl 2 , 1 mM EGTA, 1 mM DTT, 0.1% Triton X-100, 0.5 mg/ml BSA)
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Additionally, if gut permeability is indeed increased in ME/CFS patients, microbes and antigens may be able to cross the epithelial barrier into surrounding tissue and blood, potentially crossing the BBB and altering the HPA axis (443)
E., Ziegler, M
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