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A tetrasubstituted 29-amino acid peptide analog of Growth Hormone Releasing Hormone (GHRH)

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For IRE1α cleavage assay, purified RNA substrates were first incubated with inhibitor or vehicle (DMSO) for 30 min on ice, then incubated with recombinant GST-tagged human IRE1α (aa 465-977 corresponding to the kinase/ribonuclease domain, 11905-H20B, Sino Biological) in reaction buffer (40 mM HEPES pH 7.0, 10 mM Mg(OAc) 2 , 50 mM KOAc, 5 mM DTT, 1 μM ADP) for 30 minutes at 30 °C