The mRNA levels of Ly6G+ cells from the bone marrow of septic mice after IFN-gamma stimulation.
On postoperative day 5, bone marrow cells were isolated from cecal ligation and puncture (CLP) mice and sham-operated mice. A portion of the cells was immediately subjected to magnetic bead-based separation to isolate Ly6G+ cells, which were designated as the CLP group and Sham group, respectively. The remaining bone marrow cells were cultured in complete RPMI 1640 medium and stimulated with either 20 ng/mL IFN-gamma or an equal volume of PBS for 72 hours. After stimulation, Ly6G+ cells were isolated from each group using the same magnetic bead-based method. Based on IFN-gamma treatment, cells derived from CLP mice were designated as the CLP-IFN-gamma group and CLP-Control group, while those derived from sham-operated mice were designated as the Sham-IFN-gamma group and Sham-Control group. Total RNA was extracted from the Ly6G+ cells of all six groups, and RNA sequencing (RNA-seq) was performed to obtain transcriptomic data.