title: GSM8953362: TGFb-LCC-TRL 1; Homo sapiens; RNA-Seq
library_construction_protocol: Single-cell suspensions from each treatment condition were harvested and incubated with anti-human Hashtag antibodies conjugated to unique DNA barcodes and 1:100 human TruStain FcX (Fc Receptor Blocking Solution) to prevent nonspecific binding. After three PBS washes, cells from different treatment conditions were pooled for single-cell droplet encapsulation and sequencing. In both samples, Hashtag TotalSeq-A0251 and A0252 were used together to label cells following TGF-β treatment at 0 h (control), while A0253, A0254, A0255, A0256, and A0257 were used for the 3 h, 1 d, 3 d, 5 d, and 7 d time points, respectively. The original treatment conditions of individual cells were then identified based on sequencing reads of the distinct Hashtag sequences. FASTQ files of sequenced samples were processed using the SEQC pipeline  (version 0.2.1) with default parameters for the 10x single-cell 3' library and mapped to the hg38 reference human genome. The SEQC pipeline performed read alignment, multi-mapping read resolution, and cell barcode and UMI correction to generate a raw UMI-based (cell × gene) count matrix.