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

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: (A) Volcano plots depicting genes meeting fold change and p -value thresholds in MCSF and MCSF+RANKL treated cells. Differentials with −log10 transformed adjusted p -values of six and greater are plotted at the same level. Full volcano plots are included in . (B) MCSF-specific, MCSF+RANKL-specific, and common pathways altered during osteoclastogenesis. Pathways are listed in .

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: Transformation Assay