title: GSM9677623: Knockout_1; Mus musculus; RNA-Seq
library_construction_protocol: For the isolation of mBMDMs, bone marrow cells were harvested from the femurs of 8-week-old WT mice. Red blood cells were lysed using ACK lysis buffer (Beyotime, C3702, China), and the remaining cells were centrifuged at 300 × g for 5 min. The cell pellet was resuspended in α-Minimum Essential Medium (α-MEM, Gibco, C12571500BT, USA) containing 10% heat-inactivated FBS, 1% PS, and 30 ng/mL mouse macrophage colony-stimulating factor (M-CSF, MCE, HYP70553, USA). To induce macrophage polarization, cells were treated with 100 ng/mL lipopolysaccharide (LPS, Beyotime, S1732, China) for 24 h to induce inflammatory polarization.Total RNA was isolated from CD74+/+ and CD74-/- mBMDMs pre-stimulated with LPS. RNA quality and integrity were evaluated using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) High-throughput RNA sequencing libraries were prepared and sequenced on the Illumina NovaSeq 6000 platform (Illumina, San Diego, CA, USA), enabling the acquisition of high-resolution transcriptomic datasets.