title: GSM9605584: Naive-PBS-B2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA Extraction and Transcriptome Sequencing Total RNA was extracted from isolated and purified CD14⁺ monocytes using TRIzol reagent. Following ribosomal RNA (rRNA) depletion, sequencing libraries were constructed according to the Illumina library preparation protocol and subsequently sequenced on the Illumina Genome Analyzer II platform. ATAC‑seq A total of 1×10⁵ fresh cells were used for each assay. Libraries were constructed using Tn5 transposase treatment and sequenced on an Illumina platform. Raw reads were subjected to quality control using fastp and aligned to the reference genome with Bowtie2. PCR duplicates were removed using Picard, and MACS2 was used for peak calling to identify open chromatin regions (ATAC‑seq peaks). Identified peaks were annotated with HOMER, followed by transcription factor binding motif analysis. Chromatin Immunoprecipitation (ChIP) Log-phase cells were washed with PBS, cross-linked with 1% formaldehyde at room temperature for 10 min, and quenched with glycine. After lysis, chromatin was sonicated to 200–500 bp fragments. Chromatin was incubated with anti-RNA polymerase II antibody or control IgG pre-bound to Protein A/G magnetic beads at 4℃ overnight. After washing, elution, cross-link reversal and proteinase K digestion, DNA was purified and analyzed by qPCR for target region enrichment.