GSM8111769: liver, WT1, CCL4, replicate1, scRNA-seq; Mus musculus; RNA-Seq
Single-cell RNA-seq was performed following the protocol established by Gene Denovo Biotechnology Co. (Guangzhou, China).9 In brief, the cell suspension derived from mouse liver was centrifuged at 50 g for 2 min, resulting in the separation of hepatocytes settling at the bottom, while nonparenchymal cells (NPCs) were mainly remained in the supernatant suspension.9 A small amount of single cell suspension was mixed with equal volume of 0.4% trypan blue staining solution and counted using Counter ® II Automated Cell Counter. The mixture, including gel beads with barcode, cells, and enzymes, was separated by oil in the reservoir to form GEMs (Gel Beads In Emulations). Subsequently, gel beads dissolved, releasing the captured sequence containing Barcode sequence. RNA was reverse transcribed to produce cDNA, following the 10×Genomics single cell 3' library construction V3 protocol. Quantification and pooling were carried out using the ABI One-Step Plus Real-Time PCR System (Life Technologies). 9 High-throughput sequencing was performed in PE150 sequencing mode on the Illumina sequencing platform.