GSM9065913: Liver metastases from control (shSCR) 4T1 cells; Mus musculus; RNA-Seq
Following mice euthanasia, livers were perfused through the left ventricle for full body perfusion, extracted, and briefly washed in blood bank saline. Liver metastases from mice injected with 4T1 shSCR (n = 3) and 4T1 shZdhhc17 (n = 3) were then dissected under the microscope, and subsequently incubated at 37 °C for 45 min with a solution of 0.3 mg/ml liberase (Roche) and 1 µg/ml DNase1 in 2 ml of DMEM/F12 supplemented with 5% fetal bovine serum. The dissociated metastases were quenched with FACS buffer (3% fetal bovine serum in PBS supplemented with 2 mM EDTA), filtered through a 70-µm cell strainer, and centrifuged for 5 min at 300g. The cell pellets were then washed, centrifuged for 5 min at 300g, incubated with ACK lysis buffer (Merck) and strained with a 40 µm cell strainer. Single-cell suspensions were then counted, and the same number of cells were pooled together from the 3 independent liver-metastases dissociations performed per group (shSCR or shZdhhc17). The cells were then resuspended in cell culture medium and loaded on a BD Rhapsody cartridge with a targeted capture of 15,000 cells per group. Reverse transcription, cDNA amplification, and library construction were performed following the manufacturer's instructions (23-24117(02)).
GEO
GSM9065913_r1