Journal: Frontiers in Endocrinology

Article Title: Anoctamin 5 mutation leads to abnormal bone homeostasis of GDD by regulating AMPK-dependent glucose metabolism

doi: 10.3389/fendo.2026.1703491

Figure Lengend Snippet: AMPK inhibitor regulates osteogenesis and osteoclastogenesis of GDD in vivo . 12-week-old male mice were used to observe bone phenotype. (A) Representative photomicrographs and quantitative immuno-positive area analysis of OCN in tibia (bar=100 μm); (B–D) ELISA analysis of serum level of ALP, PINP (B) , OPG, OPG/RANKL (C) , and CTX (D, E) Representative images of TRAP staining and quantitative analysis of the osteoclast number (Oc.N)/perimeter of bone (B.Pm) in the germinal center of the tibia (bar=100 μm). Data are presented as mean ± SEM. Statistic significances are determined by one-way ANOVAs with Dunnett’s multiple comparison tests, with ns: no significance, *P < 0.05, **P < 0.01.

Article Snippet: Mini sample mouse receptor activator of nuclear factor kappa B ligand (RANKL) ELISA Kit (Elabscience, E-MSEL-M0084) and mouse osteoprotegerin (OPG) ELISA Kit (Elabscience, E-EL-M0081) was used to detect serum RANKL and OPG level.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Comparison

Journal: Molecular Medicine Reports

Article Title: Effects of zinc carnosine on bone loss in mice with diabetic osteoporosis

doi: 10.3892/mmr.2025.13723

Figure Lengend Snippet: COL-I, OCN, OPG and RANKL protein expression levels in each group after 8 weeks of intervention. (A) COL-I, OCN, OPG and RANKL protein expression level of each treatment group. Average optical density of (B) COL-I, (C) OCN, (D) OPG and (E) RANKL in each group. *P<0.05 vs. Ctrl group; # P<0.05 vs. T2DM model group. COL-I, type I collagen; OCN, osteocalcin; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor-κB ligand; Ctrl, control; T2DM, type 2 diabetes mellitus; ZnC, zinc carnosine; AOD, average optical density value.

Article Snippet: The primary antibodies used and their dilution ratios are as follows: Type I collagen (COL-I; 1:200; cat. no. AF7001; Affinity Biosciences), osteocalcin (OCN, 1:200; cat. no. 16157-1-AP; Proteintech Group, Inc.) and OPG (1:200; cat. no 31766-1-AP; Proteintech Group, Inc.).

Techniques: Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Effects of zinc carnosine on bone loss in mice with diabetic osteoporosis

doi: 10.3892/mmr.2025.13723

Figure Lengend Snippet: mRNA expression levels of bone metabolism-related factors. *P<0.05 and ***P<0.01 vs. Ctrl group; ## P<0.01 vs. T2DM model group. (A) Ctrl, control; (B) T2DM, type 2 diabetes mellitus; (C) ZnC, zinc carnosine. OCN, osteocalcin; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor-κB ligand.

Article Snippet: The primary antibodies used and their dilution ratios are as follows: Type I collagen (COL-I; 1:200; cat. no. AF7001; Affinity Biosciences), osteocalcin (OCN, 1:200; cat. no. 16157-1-AP; Proteintech Group, Inc.) and OPG (1:200; cat. no 31766-1-AP; Proteintech Group, Inc.).

Techniques: Expressing, Control

Journal: Stem Cell Research & Therapy

Article Title: Depletion of mmu-miR-185 enhances osteocyte connectivity and suppresses bone fragility through the interaction between ARL8B and RAB5A

doi: 10.1186/s13287-025-04612-y

Figure Lengend Snippet: Depletion of miR-185 in MLO-Y4 cells reduces the osteoclastogenic potential of RAW264.7 cells. ( A ) The levels of soluble RANKL (sRANKL) and osteoprotegerin (OPG) in the serum of WT and miR-185 KO mice were quantified by ELISA ( n = 3). ( B ) The concentrations of sRANKL and OPG in conditioned medium from WT and miR-185 KO MLO-Y4 cells were measured by ELISA ( n = 3). ( C ) RAW264.7 cells were co-cultured for 4 days with conditioned medium derived from miR-185 KO, sh Arl8b , and corresponding control MLO-Y4 cells, supplemented with 50 ng/mL RANKL and 30 ng/mL M-CSF ( n = 3). After 4 days of culture, qPCR was performed to evaluate mRNA expression levels of osteoclast marker genes, including Trap , DC-stamp , Ctsk , and Nfatc1 . ( D ) Representative images of tartrate-resistant acid phosphatase (TRAP)-stained osteoclast cultures ( n = 3). ( E ) TRAP-positive cells were counted based on the number of nuclei. ( F ) TRAP-positive multinucleated cells (containing > 3 nuclei) were quantified. ( G ) Cathepsin K (CTSK), a marker of osteoclasts, was detected by western blotting ( n = 3). ( H ) The mRNA expression of matrix metalloproteinase 14 ( Mmp14 ) in MLO-Y4 cells was measured by RT-qPCR ( n = 3). ( I , J ) The protein levels of MMP14 in MLO-Y4 cells were evaluated by western blotting. ( K ) Quantitative analysis of MMP14 protein expression was performed using ImageJ software. ( L )The mRNA expression of Mmp14 in the femurs of mice was assessed by qPCR ( n = 3). Data represent at least three independent experiments and are shown as mean ± S.D. (ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001). The full-length blot is presented in supplementary Raw Western

Article Snippet: RANKL and OPG protein levels in the supernatants were determined using the sRANKL ELISA Kit (CUSABIO, cat# CSB-E05127m) and the OPG ELISA Kit (CUSABIO, cat# CSB-E04693m), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Derivative Assay, Control, Expressing, Marker, Staining, Western Blot, Quantitative RT-PCR, Software