Sequencing Analysis of Human Adipose Derived Stem Cells Under High Glucose Conditions

Total RNA was extracted from human adipose-derived stem cells treated under high and low glucose conditions for 48 hours using the TRIzol reagent. Each group included three cell samples. The concentration and purity of the extracted RNA were assessed using a NanoDrop spectrophotometer, and RNA integrity was verified using an Agilent 2100 Bioanalyzer. Only samples with an RNA integrity number greater than 7.0 were used for subsequent RNA sequencing. RNA sequencing libraries were prepared from 1 ug of total RNA per sample using the NEBNext Ultra II RNA Library Prep Kit for Illumina per the manufacturer's guidelines. Sequencing was performed on an Illumina NovaSeq 6000 platform to produce 150 bp paired-end reads. Raw sequencing data underwent quality control checks using FastQC. Sequencing reads were aligned to the human reference genome using HISAT2, and gene expression levels were quantified and normalized to counts per million using FeatureCounts.