mouse p1np  (Elabscience Biotechnology)

Journal: Bone Research

Article Title: Spatially resolved osteoblast-traced transcriptomics uncovers TGF-β as a combination target with sclerostin in osteoporosis

doi: 10.1038/s41413-026-00521-9

Figure Lengend Snippet: Lineage tracing reveals that TGF-β regulates osteoblast-BLC conversion and reactivation. a Experimental timeline of tamoxifen pulse and TGF-β/TGFβ-Ab administration in Dmp1 -CreERt2:mTmG mice. b Representative confocal images of femoral periosteum showing GFP (green), tdTomato (red) signals, and DAPI (blue). Scale bar = 20 μm. c Violin plot of GFP+ cell thickness. Each point represents individual cells. The Wilcoxon rank-sum test was used for statistics. d Bar plot of GFP+ cells per unit length. Cell numbers were determined from three comparable sections per mouse, with eight fields (400× magnification) analyzed per section. The Wilcoxon rank-sum test was used for statistics. e Box plot of serum P1NP levels. A t -test was used for statistics. Group sizes: 8 weeks ( n = 6), 9 weeks ( n = 5), TGF-β ( n = 4), Inactive (12 weeks control + 13 weeks control; n = 6), Scl-Ab+TGF-β ( n = 3), Scl-Ab ( n = 3), TGFβ-Ab ( n = 4), and Scl-Ab+TGFβ-Ab ( n = 5). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; ns not significant

Article Snippet: Serum levels of P1NP, CTx1, TRAcP, OPG, RANKL, and TGF-β1 were measured via an enzyme-linked immunosorbent assay (ELISA) performed using ELISA kit for mouse P1NP (AC-33F1, Immunodiagnostic Systems, UK), mouse CTx1 (AC-06F1, Immunodiagnostic Systems, UK), mouse TRAcP 5b (SB-TR103, Immunodiagnostic Systems, UK), mouse OPG (MOP00, R&D systems, USA), mouse RANKL (MTR00, R&D systems, USA), and mouse TGF-β1 (DY1679-05 and DY007B, R&D systems, USA) according to the manufacturer’s instruction.

Techniques: Control

Journal: Bone Research

Article Title: Spatially resolved osteoblast-traced transcriptomics uncovers TGF-β as a combination target with sclerostin in osteoporosis

doi: 10.1038/s41413-026-00521-9

Figure Lengend Snippet: Dual inhibition of TGF-β and sclerostin increases bone mass in a hindlimb unloading model. a Experimental timeline of hindlimb unloading using C57B1/6 J mice and treatment with TGFβ-Ab, Scl-Ab, or both. Mice euthanized at week 12. b Representative μCT images of femoral diaphysis. c Representative confocal images of trabecular bone showing calcein (green), alizarin (red) signals, and DAPI (blue). Scale bar = 100 μm. d Representative images of TRAP-stained femur sections; violet = TRAP-positive, green = counterstain. Scale bar = 200 μm. e – h Bar plot of trabecular bone volume/total volume (Tra. BV/TV, e ), trabecular thickness (Tb. Th, f ), trabecular separation (Tb. Sp, g ), and trabecular number (Tb. N, h ) from μCT. Data = mean ± standard deviation. i , j Bar plot of mineral apposition rate (Tra. MAR, i ) and trabecular bone formation rate per bone surface (Tra. BFR/BS, j ) from labeling images. Data = mean ± standard deviation k Box plot of TRAP+ osteoclasts per bone surface (OC.N/BS), measured through ImageJ from TRAP-stained slides. l – n Box plot of serum P1NP ( l ) and TRAP ( m ) levels, and RANKL/OPG ratio ( n ). Group sizes: Control ( n = 5), Un_con ( n = 5), Scl-Ab ( n = 6), TGFβ-Ab ( n = 6), and Scl-Ab+TGFβ-Ab ( n = 7). Each point represents an individual mouse. ANOVA was used for statistics; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; ns not significant

Article Snippet: Serum levels of P1NP, CTx1, TRAcP, OPG, RANKL, and TGF-β1 were measured via an enzyme-linked immunosorbent assay (ELISA) performed using ELISA kit for mouse P1NP (AC-33F1, Immunodiagnostic Systems, UK), mouse CTx1 (AC-06F1, Immunodiagnostic Systems, UK), mouse TRAcP 5b (SB-TR103, Immunodiagnostic Systems, UK), mouse OPG (MOP00, R&D systems, USA), mouse RANKL (MTR00, R&D systems, USA), and mouse TGF-β1 (DY1679-05 and DY007B, R&D systems, USA) according to the manufacturer’s instruction.

Techniques: Inhibition, Staining, Standard Deviation, Labeling, Control

Journal: Calcified Tissue International

Article Title: Cxcr2 is Required for Osteoclast Regulation, Bone Structure, and Hematological Response During Bone (Re)modeling

doi: 10.1007/s00223-025-01470-x

Figure Lengend Snippet: Cxcr2 deletion expands the detected pro-inflammatory cytokines and alters systemic bone turnover. a Log-2 transformed fold change in cytokine expression (KO/WT) in Cxcr2 WT versus KO serum, quantified by Eve Technologies 32-Plex Cytokine Array. N = 6 mice per genotype. b Serum concentration of G-CSF and CXCL5 in Cxcr2 WT and KO mice. N = 3–8 mice per genotype. c Serum concentration of osteoblast and osteoclast activity markers P1NP and CTX-1 in Cxcr2 WT and KO mice. N = 4–5 mice per genotype. d Concentration of RANKL and OPG and OPG/RANKL ratio in Cxcr2 WT and KO bone conditioned media. N = 8 samples per group. Error bars represent standard deviation. Analysis of Cxcr2 WT versus KO using Welch’s t- test. Significance levels for differences are indicated: * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant ( p ≥ 0.05). N = 4–6 mice per group

Article Snippet: Levels of CTX-1 and P1NP were measured using the Serum Crosslaps (CTX-1) ELISA (Immunodiagnostic Systems #AC-02F1) and Rat/Mouse P1NP ELISA (Immunodiagnostic Systems #AC-33F1) according to the manufacturer’s protocols.

Techniques: Transformation Assay, Expressing, Concentration Assay, Activity Assay, Standard Deviation

Journal: Aging Cell

Article Title: Senescent Cells Involved in Deterioration of Bone Microstructure by High‐Frequency Parathyroid Hormone 1–34 Administration and Bone Loss

doi: 10.1111/acel.70331

Figure Lengend Snippet: Changes in bone metabolism markers, distribution of osteoclasts, and accumulation of senescent cells in response to parathyroid hormone administration. Serum bone metabolism markers ( n = 4) were analyzed. (A) The bone formation marker, amino‐terminal propeptide of type I collagen (P1NP, ng/mL). (B) Bone resorption marker tartrate‐resistant acid phosphatase 5b (TRAcP5b; U/L). (C) Number of TRAP‐positive osteoclasts per bone perimeter (N. Oc/B. Pm; 1/mm) ( n = 5). (D) Distribution of TRAP‐positive osteoclasts in the spines of young and aged mice treated with different PTH administration frequencies. RT‐qPCR analysis of mRNA expression levels of p16INK4a (E) and SASP factors ( IL6 , IL1α , IL1β , Mmp3 , MMP13 , Ccl5 ) (F–K) in osteoblast and osteocyte‐enriched cells derived from the bones of aged mice ( n = 4 mice/treatment). (L) IHC for p16INK4a in the lumbar spine of aged mice (see arrows, scale bars, 500 μm; 50 μm on right). Data represent mean ± SEM (error bars). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (one‐ or two‐way ANOVA with Tukey's multiple comparisons test). PTH, parathyroid hormone; TRAP, tartrate‐resistant acid phosphatase; SEM, standard error of the mean; ANOVA, analysis of variance; IHC, immunohistochemistry; SASP, senescence‐associated secretory phenotype; p16INK4a , cyclin‐dependent kinase inhibitor 2A; IL6 , Interleukin‐6; IL1α , Interleukin‐1 alpha; IL1β , Interleukin‐1 beta; Mmp3 , Matrix metalloproteinase‐3; MMP13 , Matrix metalloproteinase‐13; Ccl5 , C‐C motif chemokine ligand 5 .

Article Snippet: P1NP (ng/mL) and TRAcP 5b (U/L) were quantified using a Rat/Mouse P1NP enzyme immunoassay kit (Immunodiagnostic Systems, Boldon, UK) and Mouse TRAcP 5b ELISA kit, respectively.

Techniques: Marker, Quantitative RT-PCR, Expressing, Derivative Assay, Immunohistochemistry

Journal: Aging Cell

Article Title: Senescent Cells Involved in Deterioration of Bone Microstructure by High‐Frequency Parathyroid Hormone 1–34 Administration and Bone Loss

doi: 10.1111/acel.70331

Figure Lengend Snippet: Effects of senolytic treatment (dasatinib + quercetin) on parathyroid hormone‐induced senescence and bone metabolic markers. PTH treatment (10 times per week) was combined with D + Q for 4 weeks in aged mice. (A) IHC for p16INK4a in the lumbar spine (see arrows). Scale bars, 50 μm (above). Double immunofluorescence staining of p16INK4a and OPN in the lumbar spine. The arrow indicates positive p16INK4a and OPN expression. Scale bars, 10 μm. (B) Percentage of p16INK4a‐positive cells among OPN‐positive cells ( n = 5). (C) Bone formation marker P1NP (ng/mL) ( n = 5). (D) The bone resorption marker, TRAcP5b (U/L) ( n = 5). (E, F) RT‐qPCR results of Tnfsf11 and Tnfrsf11b ( n = 4). Data represent mean ± SEM (error bars). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (one‐way ANOVA with Tukey's multiple comparisons test). D + Q, dasatinib + quercetin; IHC, Immunohistochemistry; SEM, standard error of the mean; ANOVA, analysis of variance; PTH, parathyroid hormone; OPN, osteopontin; P1NP, propeptide of type I collagen; Tnfsf11 , tumor necrosis factor superfamily member 11; Tnfrsf11b , tumor necrosis factor superfamily member 1b; p16INK4a , cyclin‐dependent kinase inhibitor 2A.

Article Snippet: P1NP (ng/mL) and TRAcP 5b (U/L) were quantified using a Rat/Mouse P1NP enzyme immunoassay kit (Immunodiagnostic Systems, Boldon, UK) and Mouse TRAcP 5b ELISA kit, respectively.

Techniques: Double Immunofluorescence Staining, Expressing, Marker, Quantitative RT-PCR, Immunohistochemistry

Journal: Advanced Science

Article Title: Epigenetic Targeting of Senescent Cells Prevents the Deleterious Effects of Obstructive Sleep Apnea on Growing Skeleton

doi: 10.1002/advs.202502697

Figure Lengend Snippet: CIH induces bone loss and growth retardation in young mice. Three‐week‐old male or female wild‐type (WT) mice were subjected to CIH or normoxia condition for four weeks. A–E) Representative micro‐CT images of both male and female mice were shown in (A) with quantitative analysis of BV/TV (B), Tb. N (C), Tb. Th (D) and Tb. Sp (E). F,G) The serum concentrations of P1NP and CTX of both male and female mice were measured by ELISA. General view of the femurs was shown in H), and the length of the femurs were measured in I). J,K) H&E staining was performed to show the general morphology of the femoral bone and growth plate. RZ, resting zone; PZ, proliferating zone; HZ, hypertrophic zone; GP, growth plate. The fold change of GP length, RZ length, PZ length, and HZ length was quantified in L–O). n = 5‐6 mice Data were represented as mean ± SD. p values were determined by one‐way ANOVA.

Article Snippet: N‐terminal propeptide of type I procollagen (P1NP) was measured using P1NP ELISA kit (E‐EL‐M0233, Elabscience) and β‐CTx ELISA kit (E‐EL‐M0372, Elabscience) as described by the company.

Techniques: Micro-CT, Enzyme-linked Immunosorbent Assay, Staining

Journal: Advanced Science

Article Title: Epigenetic Targeting of Senescent Cells Prevents the Deleterious Effects of Obstructive Sleep Apnea on Growing Skeleton

doi: 10.1002/advs.202502697

Figure Lengend Snippet: Elevated H3K27me3 alleviates CIH‐induced osteoprogenitor senescence and bone loss. Three‐week‐old female UTX iKO mice and WT (UTX floxed) mice were subjected to CIH or normoxia condition for four weeks. A,B) Representative co‐immunofluorescence staining of OSX and H3K27me3 in distal femur section in (A) and analysis of cell number per mm 2 tissue area (N. OSX + H3K27me3 + cells/ Ar) in (B). C,D) Representative images of SA‐βGal staining in the distal femur section in (C) and analysis of cell number per mm 2 tissue area (N. SA‐βGal + cells/ Ar) in (D). E,F) Representative co‐immunofluorescence staining of OSX and γ H2A.X in the distal femur section in (E) and analysis of cell number per mm 2 tissue area (N. OSX + γ H2A.X + cells/ Ar) in (F). G,H) Representative immunofluorescence staining of OCN in distal femur section in (G) and analysis of cell number per mm 2 tissue area (N. OCN + cells/ Ar) in (H). Representative micro‐CT images were shown in I) with quantitative analysis of BV/TV J), Tb. N K), Tb. Th L) and Tb. Sp M). N,O) The concentrations of P1NP and CTX were measured by ELISA. n = 5‐6 mice. Data were represented as mean ± SD. p ‐values were determined by one‐way ANOVA.

Article Snippet: N‐terminal propeptide of type I procollagen (P1NP) was measured using P1NP ELISA kit (E‐EL‐M0233, Elabscience) and β‐CTx ELISA kit (E‐EL‐M0372, Elabscience) as described by the company.

Techniques: Immunofluorescence, Staining, Micro-CT, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Epigenetic Targeting of Senescent Cells Prevents the Deleterious Effects of Obstructive Sleep Apnea on Growing Skeleton

doi: 10.1002/advs.202502697

Figure Lengend Snippet: GSK‐J4 rescues CIH‐impaired bone growth and mineral acquisition. Three‐week‐old female WT mice were subjected to CIH or normoxia condition for four weeks, and at the same time, treated with GSK‐J4 at 10 mg kg −1 per day or vehicle. Representative micro‐CT images were shown in A) with quantitative analysis of BV/TV B), Tb. N C), Tb. Th D) and Tb. Sp E). F,G) The concentrations of P1NP and CTX were measured by ELISA. General view of the femurs was shown in H), and the length of the femurs were measured in I). J,K) H&E staining was performed to show tissue morphology. RZ, resting zone; PZ, proliferating zone; HZ, hypertrophic zone; GP, growth plate. The fold change of GP length, RZ length, PZ length, and HZ length were quantified in L–O). n = 5–6 mice. Data were represented as mean ± SD. p ‐values were determined by one‐way ANOVA.

Article Snippet: N‐terminal propeptide of type I procollagen (P1NP) was measured using P1NP ELISA kit (E‐EL‐M0233, Elabscience) and β‐CTx ELISA kit (E‐EL‐M0372, Elabscience) as described by the company.

Techniques: Micro-CT, Enzyme-linked Immunosorbent Assay, Staining