LincRNA-EPS regulates iron metabolism and osteoclastogenesis under inflammatory micro-environment

This study aims to investigate the impact of LincRNA-EPS on osteoclast lineage differentiation. We established LincRNA-EPS gene knockout mice on the C57BL/6 background and isolated bone marrow-derived macrophages (BMDMs) from both knockout and wild-type mice. These BMDMs were induced with RANKL in vitro for 2 days to generate pre-osteoclast cells. Transcriptome sequencing was performed on these cells, followed by further analysis of differentially expressed genes to explore the effects of LincRNA-EPS deficiency on osteoclast differentiation in BMDMs.? In this study, we found that dysregulated osteoclastogenesis and iron homeostasis without lincRNA-EPS, represented by up-regulated gene Lcn2. Overall design: ??Experimental Grouping:?? KO group:?? Bone marrow-derived macrophages (BMDMs) isolated from LincRNA-EPSknockout mice (LincRNA-EPS knockout); WT group:?? BMDMs isolated from wild-type (C57BL/6) mice. Each group included three biologically independent replicates. ??Experimental Procedure:?? ??BMDM induction:??BMDMs from both groups were cultured in complete a-MEM medium supplemented with 40 ng/mL M-CSF for 24 hours to promote macrophage proliferation and priming.Osteoclast precursor differentiation:??After initial M-CSF priming, 40 ng/mL RANKL was added to the culture medium for an additional 48 hours to induce pre-osteoclast formation . ??Sample processing:??Cells were harvested and total RNA was extracted. ??Transcriptome sequencing:??RNA sequencing was performed on the Illumina NovaSeq X plus platform.