GSM8889185: control rep2; Mus musculus; RNA-Seq
Resuspend the isolated bone marrow cells in bone marrow-derived macrophages (BMDM) growth medium (DMEM+1-%FBS+ 15% filtered (0.2 μm) L-929 cell (ATCC, CCL-1) culture supernatant) at a concentration of 2x106 cells/ml. Seed the cells in 15cm dish. Change fresh BMDM growth medium on day 3. On day 7, assess the formation of mature BMDM using flow cytometry analysis and fluorophore-conjugated antibodies to detect cells expressing CD11b and F4/80. RNA extraction was performed using FastPure Cell/Tissue Total RNA Isolation Kit V2 (Vazyme RC112-01). Reverse transcription was performed using SMARTScribe Reverse Transcriptase (Clontech) according to the manufacturer's instructions. The cDNA was amplified and the final library was constructed using TD501 kit (Vazyme).