GSM9084162: shYT; Mus musculus; RNA-Seq
Tumor-bearing livers were minced and digested in 15 mL of 1× PBS containing 7.5 mg Pronase, 7.5 mg Collagenase P, and 1.5 mg DNase I using a C Tube (Miltenyi Biotec) and the gentleMACS Octo Dissociator (Miltenyi) for 45 minutes at 100 rpm and 37 °C. Following digestion, the cell suspension was filtered through a 100 µm cell strainer and washed with an appropriate volume of Advanced DMEM/F12 (AdDMEM/F12) medium. Red blood cell lysis was performed using the Red Blood Cell Lysis Solution (Miltenyi, #130-094-183), followed by debris removal (Miltenyi, #130-109-398) and dead cell removal (Miltenyi, #130-090-101) according to the manufacturer's protocols. Cells were then incubated with Fc receptor blocking reagent (Miltenyi) for 10 minutes on ice. Subsequently, cells were labeled with 1 µg of anti-mouse hashtag antibodies (TotalSeq™-B, BioLegend) for 30 minutes on ice. After labeling, cells were washed twice with PBS and resuspended in 1× PBS. The final cell concentration was adjusted to 1,000 cells/µL for downstream single-cell library preparation. Single-cell encapsulation and barcoding were performed using the 10x Genomics Chromium Controller. The shRen includes four replicates from four different mice using the following hashtags: TotalSeq-B0301, TotalSeq-B0302, TotalSeq-B0303, TotalSeq-B0304. The shYT samples includes 4 samples: 2x shYT#1 (TotalSeq-B0301, TotalSeq-B0302) and 2x shYT#2 (TotalSeq-B0303, TotalSeq-B0304). Libraries were prepared using Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10x Genomics), according to the manufacturer's protocol.
GEO
GSM9084162_r1