β-Hydroxybutyrate alleviates intestinal aGVHD via remodeling gut microbiota-bile acid axis and inhibiting TLR4-NF-κB-driven Th1/Th17 differentiation

Published: 29 September 2026| Version 1 | DOI: 10.17632/bpzsncgwh6.1
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RNA sequencing was performed on colon tissue samples, with three biological replicates per group. Total RNA was extracted using TRIzol reagent (Invitrogen, 15596026CN), and eukaryotic mRNA was enriched using oligo(dT) magnetic beads (Beyotime, R0075) for cDNA synthesis. The resulting cDNA was fragmented, purified using the QiaQuick PCR Purification Kit (Qiagen, 28104), and subjected to Illumina library preparation (Illumina DNA Prep kit). This process included end repair, A-tailing, adapter ligation, and size selection for fragments of 300–400 bp. The libraries were sequenced on an Illumina NovaSeq 6000 platform (Metware, Wuhan, China) to generate 150‑bp paired‑end reads.

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