Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 1. (A-C) Changes in terms of grip strength, thermal pain threshold and mechanical pain threshold in three groups of mice before and after 10 weeks of intervention (D-I) Levels of serum IFN-γ, IL-10, RANKL, IL-17 A, TNF-α, and TGF-β after 10 weeks of behavioural monitoring. n = 12, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques:

Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 2. (A-B) Changes in bone metabolism biomarkers, bone microstructure and related protein expression in three groups of mice after CBT intervention. (C-D) Representative images of ABH staining (40× scale bar: 1000 μm, 100× scale bar: 400 μm) (E) Analysis of the distal femoral epiphyseal region of mice using micro- computed tomography (CT). (F-G) Representative images of Immunohistochemistry of RANKL and OPG. (H-K) Comparison of BV/TV, Tb.Th, Tb.N, Tb.Sp. (L) Protein blot analysis of bone tissue targeting proteins associated with the RANKL/OPGpathway and inflammatory factors. (M-P) Expression of RANKL, TNF-α, OPG, and TGF-β1 proteins in bone tissues. n = 12, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques: Expressing, Staining, Micro-CT, Immunohistochemistry, Comparison

Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 6. (A-C) Spearman’s correlation analysis of gut microbiota at phylum, family and genus level with serum factors IFN-γ, IL-10, RANKL, IL-17 A, TNF-α and, TGF-β.(D) PICRUSt2 analysis identifies the top 10 KEGG pathways.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques:

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Implant wear induces inflammation, but not osteoclastic bone resorption, in RANK(-/-) mice.

doi: 10.1002/jor.20190

Figure Lengend Snippet: Figure 3. Macrophage staining and immunohistological staining of CD68 and RANKL in pouch membrane. The number of infiltrating macrophages in pouch membrane was evaluated by both immunostaining of CD 68, a macrophage cell marker (A) and nonspecific esterase (NSE) histological stain (B). Immunohistological staining of RANKL in pouch membrane was shown in (C). B, implanted bone; M, pouch membrane. Arrows indicate positive staining. (Original magnification, 200).

Article Snippet: Sections were blocked with 1.5% normal goat serum for 1 h, and incubated with goat anti-mouse TNFa, IL-1b, CD68, and RANKL antibodies (2 mg/ml, Santa Cruz Biotechnology, Santa Cruz, CA) at 48C overnight.

Techniques: Staining, Membrane, Immunostaining, Marker

Journal: Journal of Dental Sciences

Article Title: T cell and periosteum cooperation in osteoclastogenesis induced by lipopolysaccharide injection in transplanted mouse tibia

doi: 10.1016/j.jds.2018.03.001

Figure Lengend Snippet: (A) The information of animal experimental schedule (date), groups, and sample size(n). (B) Schema of the tibial surface for histometric analysis. The number of TRAP-positive mononuclear and multinuclear cells, RANKL-positive cells, and CD3-positive cells was counted in a rectangular area (50 × 1000 μm) on the center of the tibia.

Article Snippet: Sections were then immersed in rabbit anti-mouse RANKL polyclonal antibody (1:200; Acris Antibodies, Rockville, MD, USA) and CD3 polyclonal antibody (1:200; Abcam, Cambridge, UK) at 4 °C overnight.

Techniques:

Journal: Journal of Dental Sciences

Article Title: T cell and periosteum cooperation in osteoclastogenesis induced by lipopolysaccharide injection in transplanted mouse tibia

doi: 10.1016/j.jds.2018.03.001

Figure Lengend Snippet: Immunohistological findings (A–D), and the number of RANKL-positive cells (E). (A–D) Arrowheads indicate RANKL-positive cells. (E) The number of RANKL-positive cells was highest in the WT (+) LPS group. Scale bars represent 25 μm. Bars represent mean ± SD. * P < 0.01 vs. WT (+) LPS group.

Article Snippet: Sections were then immersed in rabbit anti-mouse RANKL polyclonal antibody (1:200; Acris Antibodies, Rockville, MD, USA) and CD3 polyclonal antibody (1:200; Abcam, Cambridge, UK) at 4 °C overnight.

Techniques:

Journal: Journal of cellular physiology

Article Title: Autoregulation of RANK ligand in oral squamous cell carcinoma tumor cells

doi: 10.1002/jcp.26456

Figure Lengend Snippet: Effect of RANKL autoregulation in OSCC cell on osteoclast differentiation/bone resorption. (a) RANKL CRISPR-cas9 knockout (KO) plasmid transfected SCC14a cells as visualized by GFP expression under fluorescence microscopy. (b-i) Western blot analysis of RANKL expression in RANKL-KO OSCC cells. β-actin expression served as control. (b-ii) Total RNA isolated from OSCC cells transfected with CRISPR-Cas9 double nickase plasmid and subjected to real-time RT-PCR analysis of RANKL mRNA expression. Relative mRNA expression level was normalized with respect to GAPDH amplification. The values are expressed as mean ± SD (*p < 0.05). (c and c-i) OSCC-CM effect on osteoclast differentiation. Mouse bone marrow non-adherent cells were incubated with M-CSF (10 ng/ml), RANKL (50 ng/ml) with or without normal and RANKL-KO OSCC-CM (20%) for 7 days. Cells cultured with M-CSF alone served as control. TRAP-positive multinucleated osteoclasts formed at the end of the culture period were scored. The values are expressed as mean ± SD (*p < 0.05). *Compared to control cultures; #Compared to normal OSCC-CM treated cells. (d) The mouse bone marrow non-adherent cells were cultured as above on dentine slices for 10 days in the presence of M-CSF (10 ng/ml), RANKL (50 ng/ml) with or without normal and RANKL-KO OSCC-CM (20%). (d-i) The resorbed area of dentine slices was calculated relative to the total area. (d-ii) The osteoclasts formed on the surface of dentine slices were counted under light microscope. Each bar represents the mean ± SD (p < 0.05). *Compared to control cultures; #Compared to normal OSCC-CM treated cells

Article Snippet: We used CRISPR double nickase plasmid (Santa Cruz) to suppress RANKL gene expression in OSCC tumor cells.

Techniques: CRISPR, Knock-Out, Plasmid Preparation, Transfection, Expressing, Fluorescence, Microscopy, Western Blot, Control, Isolation, Quantitative RT-PCR, Amplification, Incubation, Cell Culture, Light Microscopy

Journal: Drug Design, Development and Therapy

Article Title: Sinomenine Inhibits Orthodontic Tooth Movement and Root Resorption in Rats and Enhances Osteogenic Differentiation of PDLSCs

doi: 10.2147/DDDT.S379468

Figure Lengend Snippet: Results of immunohistochemical staining. ( A – B ) The result of TNF-α ( A ) and RANKL ( B ) immunostaining of the maxillary first molar indicated that the expression of TNF-α and RANKL decreased in sinomenine-treated groups. ( C – E ) The results of OPG ( C ), RUNX2 ( D ) and OCN ( E ) IHC staining indicated that the expression of OPG, RUNX2 and OCN increased. Scale Bar: 20 μm. Data are expressed as mean ± SD (* P<0.05, **P<0.01, ***P<0.001).

Article Snippet: The blots were probed with primary antibodies overnight at 4 °C to ALP (Huabio, Hangzhou, China), RUNX2 (Proteintech, Hubei, China), OPG (Abcam, Cambridge, UK), RANKL and GAPDH (Proteintech, Hubei, China).

Techniques: Immunohistochemical staining, Staining, Immunostaining, Expressing, Immunohistochemistry

Journal: Drug Design, Development and Therapy

Article Title: Sinomenine Inhibits Orthodontic Tooth Movement and Root Resorption in Rats and Enhances Osteogenic Differentiation of PDLSCs

doi: 10.2147/DDDT.S379468

Figure Lengend Snippet: Effect of sinomenine on proliferation and osteogenesis of PDLSCs. ( A ) PDLSCs were treated with 0, 0.1 M, 0.2 M, 0.5 M, 1.0 M, 2.0 M sinomenine for 0, 1, 3, and 5 days, and the growth curves were depicted according to the results of CCK-8 assay. ( B ) 0.1 M sinomenine stimulated the proliferation of PDLSCs, while 1.0 M and 2.0 M sinomenine markedly inhibited the proliferation of PDLSCs on day 5. ( C ) Mineral deposition was observed by Alizarin red staining. Scale bar: 200 μm. ( D ) The relative amount of mineralized nodule was assessed based on the absorbance at 562 nm. ( E ) ALP activity was enhanced by 0.1 M and 0.5 M sinomenine at day 7. ( F and G ) The mRNA and protein expression levels of ALP ( F ) and RUNX2 ( G ) in sinomenine-treated groups for 7 days. Data are presented as mean ± SD (**P<0.01, ***P<0.001, ****P<0.0001).

Article Snippet: The blots were probed with primary antibodies overnight at 4 °C to ALP (Huabio, Hangzhou, China), RUNX2 (Proteintech, Hubei, China), OPG (Abcam, Cambridge, UK), RANKL and GAPDH (Proteintech, Hubei, China).

Techniques: CCK-8 Assay, Staining, Activity Assay, Expressing

Journal: Oncotarget

Article Title: Luminal STAT5 mediates H2AX promoter activity in distinct population of basal mammary epithelial cells

doi: 10.18632/oncotarget.9718

Figure Lengend Snippet: ( A ) The effect of 5-aza on the number of visible H2AX–GFP clones (≥ 1 cell) was monitored in BLG-STAT5/H2AX–GFP-transfected cultures on day 4 of treatment, in the presence or absence of PRL and hydrocortisone. Bars represent the number of visible H2AX–GFP cells/clones (GFP + ) per 100,000 seeded cells. The clones were monitored in 108–204 wells (96-well plate) seeded with 1000 cells each. Chi square values were calculated and significant differences ( P < 0.001), indicated by an asterisk, were determined using Pearson test. Inset: two GFP-expressing clones. Bar = 50 μm. ( B ) The effect of RANKL siRNA transfection on RANKL expression was determined by real-time PCR after 3 days of treatment with PRL, hydrocortisone and 5-aza. Bars represent an average of two independent transfections with range determination of RANKL, or non-specific siRNA. ( C ) RANKL siRNA transfection suppressed the number of GFP + cells/clones under the conditions elaborated in (B). Bars represent the number of GFP + clones per 50,000 seeded cells, monitored in 28 or 48 wells, seeded with 2000 cells each and monitored on day 4 of culture. Chi square values were calculated and significant differences ( P < 0.01), indicated by an asterisk, were determined using Pearson test.

Article Snippet: Transient transfection of RANKL siRNA (mouse, Santa Cruz Biotechnology, Dallas, TX) was performed according to the manufacturer's protocol and cells were supplemented for 3 days with DMEM:F12 containing FCS, insulin, hydrocortisone, PRL and 5-aza.

Techniques: Clone Assay, Transfection, Expressing, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Luminal STAT5 mediates H2AX promoter activity in distinct population of basal mammary epithelial cells

doi: 10.18632/oncotarget.9718

Figure Lengend Snippet: Paracrine signaling via RANKL (and possibly other transducers) from rare luminal cells with highly activated STAT5 induces demethylation and activation of the H2AX promoter in their neighboring basal cells. This includes the involvement of yet undetermined transcription factors. Note: a direct effect of STAT5 overexpression in a single luminal cell on RANKL secretion, and the mechanism by which RANKL affects demethylation of the H2AX promoter in neighboring basal cells have yet to be demonstrated.

Article Snippet: Transient transfection of RANKL siRNA (mouse, Santa Cruz Biotechnology, Dallas, TX) was performed according to the manufacturer's protocol and cells were supplemented for 3 days with DMEM:F12 containing FCS, insulin, hydrocortisone, PRL and 5-aza.

Techniques: Activation Assay, Over Expression