RNA-Seq of Homo sapiens: HUVECs treated with AGEs for 48 h, biological replicate 3
Total RNA was extracted from HUVECs treated with advanced glycation end products (AGEs) for 48 h using a one-step column-based extraction method. RNA quality was assessed using a NanoDrop 2000 spectrophotometer, agarose gel electrophoresis, and an Agilent 2100 Bioanalyzer. Poly(A)-containing mRNA was enriched using oligo(dT) magnetic beads and fragmented to approximately 300 bp. First-strand cDNA was synthesized using random hexamer primers, followed by second-strand cDNA synthesis, end repair, A-tailing, adapter ligation, and 15 cycles of PCR amplification. Libraries were prepared using the Illumina TruSeq RNA Sample Preparation Kit and sequenced in paired-end mode (2  150 bp) on an Illumina NovaSeq 6000 platform.