nf kb ligand  (MedChemExpress)

Journal: ACS Applied Materials & Interfaces

Article Title: Regulation of Bone Remodeling by Metal–Phenolic Networks for the Treatment of Systemic Osteoporosis

doi: 10.1021/acsami.4c18829

Figure Lengend Snippet: In vitro osteoclastogenic differentiation and activation of RAW264.7 cells induced by t-MPN NPs. (A) Experimental timeline of RANKL-induced RAW264.7 cells cultured with t-MPN NPs on osteoclastogenic differentiation and activation. Cells treated with Sr, EGCG, and nt-MPN NPs were used for comparison. (B) TRAP staining of RAW264.7 cells cultured with different treatments for 7 days. Scale bars is 100 μm. (C) Quantitative analysis of the area (top) and viability (bottom) of TRAP-positive osteoclasts of RAW264.7 cells cultured with various treatments for 7 days. Scale bars are 50 μm. (D) F-actin (red)/nucleus (blue) staining of RAW264.7 cells cultured with different treatments on days 3 and 7. Scale bar is 50 μm. (E) Quantitative analysis of the osteoclasts number per 0.065 mm 2 area at day 7. (F) Expression of osteoclastogenic genes (TRAP, CTSK, MMP9, and NFATc1) in RAW264.7 cells determined by qRT-PCR analysis for 7 days. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n.s. not significant.

Article Snippet: CY5 and receptor activator of NF-kB ligand (RANKL/TNFSF11) were purchased from MedChemExpress (Shanghai, China).

Techniques: In Vitro, Activation Assay, Cell Culture, Comparison, Staining, Expressing, Quantitative RT-PCR

Journal: JBMR Plus

Article Title: Mouse mandibular–derived osteoclast progenitors have differences in intrinsic properties compared with femoral–derived progenitors

doi: 10.1093/jbmrpl/ziae029

Figure Lengend Snippet: Monocytes isolated from mandible bone marrow have enhanced osteoclast differentiation. Bone marrow cells were flushed from mandibular and femoral bone and monocytes were selected for osteoclast cultures from 2–month–old mice. (A) Schematic illustrating procedure for selecting monocytes from bone marrow for osteoclast cultures; (B) representative TRAP–stained images of monocytes differentiated into osteoclasts in the presence of M-CSF and RANKL for 4 days. (C) Quantification of the number of TRAP positive multinuclear cells ( N ≥ 3). (D) Quantification of size of TRAP positive multinuclear cells. Scale bar is 10 mm.

Article Snippet: Two days later, cells were refed with 1.5% CMG 14-12 culture supernatant and 5 ng/mL of receptor activator of NF-kB ligand (RANKL) (R and D Systems, catalog #462-TEC-010) to stimulate osteoclast differentiation.

Techniques: Isolation, Staining

Journal: JBMR Plus

Article Title: Mouse mandibular–derived osteoclast progenitors have differences in intrinsic properties compared with femoral–derived progenitors

doi: 10.1093/jbmrpl/ziae029

Figure Lengend Snippet: Mandibular–derived osteoclast precursors proliferate less than femoral–derived osteoclast precursors and do not exhibit an increase in apoptosis activity. Bone marrow cells were flushed from the mandible and femur of 2–month–old C57Bl/6 mice and monocytes were selected. (A) Monocytes were grown in M-CSF for 4 days. CCK-8 activity was measured at 450 nm at plating (day 0) or after 4 days in M-CSF. (B) Osteoclast precursors were grown in M-CSF (day 0) or M-CSF and RANKL for 2 days (day 2). Apoptosis activity was quantified at day 0 or 2 using Promega Caspase-Glo 3/7 Assay. Graphed data represent Caspase-Glo3/7 activity normalized to the average number of nuclei in each culture condition ( N ≥ 3). Samples were compared using Student’s t -test.

Article Snippet: Two days later, cells were refed with 1.5% CMG 14-12 culture supernatant and 5 ng/mL of receptor activator of NF-kB ligand (RANKL) (R and D Systems, catalog #462-TEC-010) to stimulate osteoclast differentiation.

Techniques: Derivative Assay, Activity Assay, CCK-8 Assay, Caspase-Glo Assay