rat rankl recombinant protein  (R&D Systems)

Journal: Journal of Nanobiotechnology

Article Title: Cerium-doped prussian blue nanozymes for bone regeneration and osteoprotection by Lhx2 in sutural distraction osteogenesis through multi-omics analysis

doi: 10.1186/s12951-025-03931-9

Figure Lengend Snippet: Systematical observation and analysis of bone remodeling and regeneration within the mid-palatal suture expansion by histological and immunohistochemical staining. ( A ) HE and Masson’s trichrome staining. ( B ) TRAP staining and IHC staining of OCN, CD31, RANKL and OPG expression. Black arrows indicate osteoclasts in mid-palatal suture. ( C ) The number of osteoclasts per square millimeter was quantified through TRAP staining. H-Score of ( D ) OCN, ( E ) CD31, ( F ) RANKL and ( G ) OPG. ( H ) The relative value of H-Score of RANKL to OPG (* p < 0.05, ** p < 0.01, *** p < 0.001 and ns means no significance). H -Score: histochemistry score

Article Snippet: Rat M-CSF recombinant protein was purchased from PeproTech ® , USA and Rat RANKL recombinant protein was purchased from R&D Systems, USA.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Cerium-doped prussian blue nanozymes for bone regeneration and osteoprotection by Lhx2 in sutural distraction osteogenesis through multi-omics analysis

doi: 10.1186/s12951-025-03931-9

Figure Lengend Snippet: Systematical observation and analysis of bone remodeling and regeneration within the mid-palatal suture excessive expansion by histological and immunohistochemical staining. ( A ) HE and Masson’s trichrome staining. (B ) TRAP staining and IHC staining of OCN, CD31, RANKL and OPG expression. Black arrows indicate osteoclasts in mid-palatal suture. ( C ) The number of osteoclasts per square millimeter was quantified through TRAP staining. H-Score of ( D ) OCN, ( E ) CD31, ( F ) RANKL and (G ) OPG. ( H ) The relative value of H-Score of RANKL to OPG (* p < 0.05, ** p < 0.01, *** p < 0.001 and ns means no significance). H-Score: histochemistry score

Article Snippet: Rat M-CSF recombinant protein was purchased from PeproTech ® , USA and Rat RANKL recombinant protein was purchased from R&D Systems, USA.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Expressing

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 1 | SA inhibits RANKL-induced osteoclast differentiation and F-Actin ring formation in RAW 264.7 cells. (A) The chemical structure of SA. (B) Representative images of TRAP staining showing that SA inhibited RANKL-induced osteoclast differentiation in RAW 264.7 cells (original magnification ×40). (C) The TRAP-positive multinucleated cells with more than three nuclei were counted as mature osteoclasts under a microscope. (D) Representative images of F-Actin ring immunofluorescence staining showing that SA inhibited RANKL-induced F-Actin ring formation in RAW 264.7 cells (original magnification ×200). (E) The number of F-Actin rings/well was calculated under a fluorescence microscope. (F) SA showed no cytotoxic effect on RAW 264.7 cells, as determined by MTT assay. Data are expressed as the mean ± SEM of three independent biological experi- ments. ***p < 0.001 versus the control group; ###p < 0.001 versus the group only treated with RANKL.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Staining, Microscopy, Immunofluorescence, Fluorescence, MTT Assay, Control

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 2 | SA inhibits osteoclast-specific gene and protein expression during osteoclastogenesis. (A–E) SA inhibited RANKL-induced mRNA expression of TRAP (A), c-Src (B), cathepsin K (C), β3-Integrin (D) and MMP-9 (E) in RAW 264.7 cells, as determined by qRT-PCR analysis. (F) SA inhibited RANKL-induced protein expression of TRACP-5b, c-Src and cathepsin K in RAW 264.7 cells, as determined by western blotting analysis. Representative immunoblots are displayed. (G–I) The relative protein expression corresponding to TRACP-5b (G), c-Src (H) and cathepsin K (I) was calculated using AlphaView software. Total RNA or proteins from RAW 264.7 cells pretreated with or without SA for 20 min, then stimulated with RANKL (50 ng/mL) for 72 h, were subjected to qRT-PCR or western blotting analysis. Data are expressed as the mean ± SEM of three independent biological experiments. ***p < 0.001 versus the control group; ###p < 0.001 versus the group only treated with RANKL.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 3 | SA inhibits RANKL-induced expression of key transcription factors NFATc1 and c-Fos in RAW 264.7 cells. (A, B) SA inhibited RANKL-induced mRNA expression of NFATc1 (A) and c-Fos (B) in RAW 264.7 cells, as determined by qRT-PCR analysis. (C and D) SA inhibited RANKL-induced protein expression of NFATc1 (C) and c-Fos (D) in RAW 264.7 cells, as determined by western blotting analysis. Representative im- munoblots are displayed. The relative protein expression corresponding to NFATc1 and c-Fos was calculated using AlphaView software. Treatment conditions were the same as described in Figure 2. Data are expressed as the mean ± SEM of three independent biological experiments. ***p < 0.001 versus the control group; ##p < 0.01 and ###p < 0.001 versus the group only treated with RANKL.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software, Control

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 5 | SA blocks RANKL–RANK interaction and inhibits RANKL-induced RANK–TRAF6 association. (A and B) Direct interactions be- tween SA and RANK (A) and RANKL (B) were determined by SPR kinetic analysis. (C) SA blocked the interaction between RANKL and RANK, as determined by SPR-based “A-B-A" competition assay. The data represent one of three independent experiments with similar results. (D, E) SA suppressed RANKL-stimulated interaction between RANK and TRAF6 in RAW 264.7 cells, as validated by co-immunoprecipitation analysis. Representative immunoblots are displayed. The relative protein expression corresponding to TRAF6 was calculated using AlphaView software. Data are expressed as the mean ± SEM of three independent biological experiments. ***p < 0.001 versus the control group; ###p < 0.001 versus the group only treated with RANKL.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Competitive Binding Assay, Immunoprecipitation, Western Blot, Expressing, Software, Control

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 7 | SA prevents OVX-induced bone loss in mice by inhibiting osteoclastogenesis. (A) H&E staining (original magnification ×100) and TRAP staining (original magnification ×400) of the distal femur sections in each group. Representative images are displayed. (B) The trabecular bone area in the femur was quantified. (C) The number of TRAP-positive cells/field was calculated. (D–I) The serum levels of TRACP-5b (D), CTX-I (E), RANKL (F), OCN (G), PINP (H) and OPG (I) were determined using the corresponding ELISA kits. Data are expressed as the mean ± SEM (n = 10). *p < 0.05 and ***p < 0.001 versus the sham group; #p < 0.05 and ##p < 0.01 versus the OVX group.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal of cellular and molecular medicine

Article Title: Stephanine Protects Against Osteoporosis by Suppressing Osteoclastogenesis via Inhibition of the RANKL-RANK Interaction.

doi: 10.1111/jcmm.70256

Figure Lengend Snippet: FIGURE 9 | A schematic model of the molecular mechanisms under- lying the inhibitory effect of SA on RANKL-induced osteoclastogenesis.

Article Snippet: The recombinant mouse RANKL protein used in the cell experiments was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: