GSM7656511: Gene expression KO1; Mus musculus; RNA-Seq
Fresh liver tissues were dissected from HCC mice or control mice, minced and dispersed with scissors in 5 mL tubes, and then digested with 1mg/mL Collagenase IV at 37℃, 120rpm for 45 mins. The cell suspension was screened with 200 mesh filter, centrifuged at 450g, 5 mins, and aspirated supernatant from cell pellet. Further, the tumor-infiltrating leukocytes were purified by 40% percoll gradient centrifugation at room temperature for 20 mins with the acceleration and brake set to medium settings. For flow sorting of macrophages, cell suspension was stained with appreciated antibodies and resuspended at a concentration of 1×107 cells/mL in 1× PBS (pH 7.4) containing 0.5 mM EDTA and 2% FBS, and analyzed with BD Aria II (18 color sorter, USA). Cells were lysed with Trizol reagent (Invitrogen, USA) and used for microarray analysis with the 4 × 44 K Agilent Whole Mouse Genome Oligo Microarray (Novogene, China) or real-time quantitative PCR. RNA was isolated according to the standard protocol.