Journal: PeerJ
Article Title: RANK signaling in osteoclast precursors results in a more permissive epigenetic landscape and sexually divergent patterns of gene expression
doi: 10.7717/peerj.14814
Figure Lengend Snippet: Male and female bone marrow-derived macrophages were cultured with 25 ng/mL MCSF alone or 25 ng/mL MCSF and 100 ng/mL RANKL for 3 days. (A) Micrographs of macrophages and osteoclasts prior to lysis and RNA extraction. Cells cultured with RANKL demonstrated multinucleation and increased size. (B) Micrographs of TRAP-stained osteoclast differentiated under conditions matched to RNA source cells.
Article Snippet: Osteoclast precursors from male and female mice were divided into two treatment groups: one group of cells (macrophages) were maintained in α -MEM with 25 ng/mL MCSF for 72 h; the other group of cells (osteoclasts), were maintained in α -MEM with 25 ng/mL MCSF (200-08; Shenandoah Biotechnology, Warminster, PA, USA) and 100 ng/mL RANKL (200-04; Shenandoah Biotechnology, Warminster, PA, USA) for 72 h. Media were refreshed at 48 h. At the conclusion of the culture period, cells were lysed in TRI reagent and RNA was extracted using the Direct-zol RNA miniprep kit (R2051; Zymo Research, Irvine, CA, USA).
Techniques: Derivative Assay, Cell Culture, Lysis, RNA Extraction, Staining