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RNA was isolated from the brain using TRIzol reagent (Invitrogen, Thermo Fisher Scientific) following the manufacturer’s instructions, and treated with DNase I (Invitrogen, Thermo Fisher Scientific) to eliminate genomic DNA. Reverse transcription was performed using the iScript cDNA Synthesis Kit (Bio-Rad Laboratories). PCR for XBP1 was performed to detect XBP1s mRNA using Taq DNA Polymerase (QIAGEN). PCR products were assessed by electrophoresis on a 3% agarose gel as previously described (Stone et al., 2018, 2020). There were 3 mice in each group.
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