antibodies targeting osteocalcin  (TaKaRa)

Journal: JCI Insight

Article Title: Angiopoietin-like 8 governs osteoblast-adipocyte lineage commitment during skeletal aging

doi: 10.1172/jci.insight.189371

Figure Lengend Snippet: ( A ) Weight of the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. n = 7–8. ( B ) H&E staining the femur of the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. Scale bars: 100 μm. ( C and D ) Quantification of number ( C ) and area ( D ) of adipocytes in the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. n = 3. ( E ) Representative microCT images of distal femurs and midshaft cortical bone from the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. Scale bar: 1 mm. ( F – I ) Quantitative microCT analyses of the distal end of the femurs from the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. n = 7–8. BMD, bone mineral density; BV/TV, bone volume per tissue volume; Ct.Th, cortical bone thickness; Tb.N, trabecular number. ( J ) Representative osteocalcin-positive cell images of distal femurs from the GW9662-treated WT and Angptl8- Nestin-cre Tg male mice at 18 months. Scale bars: 50 μm. ( K ) Quantification of osteocalcin + cells on the bone surface (number of osteocalcin + cells per bone perimeter, N.Ocn + /B.Pm). Data are mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 by 2-way ANOVA followed by Tukey’s multiple-comparison test.

Article Snippet: Following antigen retrieval, bone sections underwent blocking with 5% BSA for 1 hour at room temperature and were then incubated overnight at 4°C with primary antibodies targeting osteocalcin (Servicebio, GB11233-100) or β-catenin (Servicebio, GB12015-100).

Techniques: Staining, Comparison

Journal: JCI Insight

Article Title: Angiopoietin-like 8 governs osteoblast-adipocyte lineage commitment during skeletal aging

doi: 10.1172/jci.insight.189371

Figure Lengend Snippet: ( A ) qRT-PCR analyses of the mRNA expression of Angptl8 in the BMMSCs transfected with control-ASO and Angptl8 -ASO. n = 3. ( B ) Experimental scheme to demonstrate that Angptl8 -ASO partially rescues the phenotype of aging male mice. ( C ) H&E staining of the femurs of the control-ASO and Angptl8 -ASO male mice at 18 months. Scale bar: 100 μm. n = 8. ( D and E ) Quantification of number ( D ) and area ( E ) of adipocytes in the control-ASO and Angptl8 -ASO male mice at 18 months. n = 3. ( F ) Representative microCT images of distal femurs and midshaft cortical bone from the control-ASO and Angptl8 -ASO male mice at 18 months. Scale bar: 1 mm. n = 8. ( G – J ) Quantitative microCT analyses of the distal end of the femurs from the control-ASO and Angptl8 -ASO male mice at 18 months. n = 8. BMD, bone mineral density; BV/TV, bone volume per tissue volume; Ct.Th, cortical bone thickness; Tb.Th, trabecular thickness. ( K ) Representative osteocalcin + cell images of distal femurs from the control-ASO and Angptl8 -ASO male mice at 18 months. Scale bar: 50 μm. n = 8. ( L ) Representative β-Gal staining images of distal femurs from the control-ASO and Angptl8 -ASO male mice at 18 months. Scale bar: 100 μm. n = 8. ( M ) Representative TRAP staining images of distal femurs from the control-ASO and Angptl8 -ASO male mice at 18 months. Scale bar: 100 μm. n = 8. Data are mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 by 2-tailed Student’s t test.

Article Snippet: Following antigen retrieval, bone sections underwent blocking with 5% BSA for 1 hour at room temperature and were then incubated overnight at 4°C with primary antibodies targeting osteocalcin (Servicebio, GB11233-100) or β-catenin (Servicebio, GB12015-100).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Control, Staining

Journal: Advanced Science

Article Title: Manganese Enhances the Osteogenic Effect of Silicon‐Hydroxyapatite Nanowires by Targeting T Lymphocyte Polarization

doi: 10.1002/advs.202305890

Figure Lengend Snippet: In vitro osteogenic effects and effective cytokine responses induced by Mn‐SiHANW–stimulated T helper (Th) cells. A) Alkaline phosphatase (ALP) staining and B) quantification of bone marrow stromal cells (BMSCs) incubated in conditioned medium derived from PBS (Control), SiHANWs, or 5% Mn‐SiHANWs with or without T helper cells. * P < 0.05, ** P < 0.01, *** P < 0.001. C) RT‐PCR analysis of OCN, Osteopontin (Opn), Osteonectin, and Collagen I gene expressions in BMSC stimulated by the conditioned medium. * P < 0.05, ** P < 0.01, *** P < 0.001 versus the Th+PBS group. D) IL‐4, E) IFN‐γ, and F) their ratio in conditioned medium of Th+PBS, Th+SiHANWs, or Th+5%SiHANWs. G) Reverse transcription polymerase chain reaction (RT‐PCR) analysis of anti‐ and pro‐inflammatory genes. * P < 0.05, ** P < 0.01, *** P < 0.001. H) Flow cytometry analysis of Th1 (T‐bet + ) and Th2 (Gata3 + ). I) The proportion of Th1, J) Th2, and K) their ratio of CD4 + T cells stimulated with PBS, SiHANWs, and 5% Mn‐SiHANWs. * P < 0.05, ** P < 0.01, *** P < 0.001. L) ALP staining and M) quantification of BMSC stimulated by the conditioned medium supplemented with IL‐4 recombinant protein, anti‐IL‐4 antibodies, or IFN‐γ recombinant protein. * P < 0.05, ** P < 0.01. N) RT‐PCR analysis of OCN, Opn, Osteonectin, and Collagen I gene expressions in BMSC stimulated by the conditioned medium. # P < 0.05, ## P < 0.01, ### P < 0.001 versus the Th+SiHANW group; + P < 0.05, ++ P < 0.01, +++ P < 0.001 versus the Th+5% Mn‐SiHANW group. N = 3.

Article Snippet: The application of primary antibodies targeting OCN protein (1:150, 23418‐1‐AP, proteintech, UK) ensued, and after an overnight incubation at 4 °C, sections were visualized using an HRP‐conjugated compact polymer system, with DAB serving as the chromogen.

Techniques: In Vitro, Staining, Incubation, Derivative Assay, Control, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Polymerase Chain Reaction, Flow Cytometry, Recombinant