RNA sequencing of HTR-8/SVneo cells treated with LPS or co-treated with LPS and Amuc_1100 (1)

HTR-8/SVneo cells were treated with 200 ng/mL LPS or co-treated with 200 ng/mL LPS and 1000 ng/mL Amuc_1100 for 24 h (n=3) . Total RNA was extracted using TRIzol reagent (Thermo Fisher Scientific, USA), and RNA purity and integrity were assessed using a NanoDrop 2000 spectrophotometer and Agilent 2100 Bioanalyzer (Agilent Technologies, USA), respectively.

Libraries were constructed with the TruSeq Stranded mRNA Library Prep Kit (Illumina, USA) and sequenced on the Illumina NovaSeq 6000 platform (paired-end, 150 bp). After quality control with fastp, clean reads were mapped to the human reference genome (GRCh38) using HISAT2, and gene-level quantification was performed with featureCounts.