title: GSM8755610: Gene expression of JQ1, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: MPE CD3+ T cells were treated with or without 500 nM JQ1 for 3 days (D3R3, n=3). Multiplex single-cell sequencing analysis was performed using TotalSeq™-B antibodies (394645, 394647, 394635, and 394637, BioLegend) as per the manufacturer's instructions. Briefly, 2 × 10^6 cells were resuspended in Cell Staining Buffer (BioLegend) and incubated with Human TruStain FcX™ Fc Blocking Reagent (BioLegend) for 10 minutes at 4°C. TotalSeq™-B antibodies were then added to each cell suspension and incubated for 30 minutes at 4°C. After antibody labeling, the samples were pooled, filtered through a 40 µm Flowmi™ Cell Strainer (BAH136800040, Sigma), and the cell concentration and viability were assessed using a Countess II FL Automated Cell Counter (Invitrogen™). Single-cell libraries were constructed using the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (10× Genomics, CG000206 Rev D), incorporating Feature Barcoding technology for cell surface protein analysis. To target a recovery of 10,000 cells, the cell suspension was adjusted to 700–1,200 viable cells per microliter before generating single628 cell gel beads in emulsion (GEM). Libraries were sequenced on a NovaSeq 6000 System (Illumina) using paired-end 150 bp reads, generating at least 80 GB of data per sample for gene transcriptome sequencing and 20 GB for hashing tags. Single-cell gene expression library was prepared by the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (10x Genomics) following the manufacturer's instructions.