Journal: Journal of Orthopaedic Translation

Article Title: A beneficial effect of a novel DKK1 monoclonal antibody and its sequential alendronate on bone and muscle properties of orchiectomized mice

doi: 10.1016/j.jot.2025.10.014

Figure Lengend Snippet: Staining of HE, TRACP, Von Kossa and bone mineral apposition rate of bone A. HE staining of the left tibia, scale bar = 100 um B. TRACP staining of the left tibia, scale bar = 100 um C. Von Kossa staining of the left tibia, scale bar = 1500 um D. Von Kossa staining of the L3 vertebra, scale bar = 1500 um E. Image of cortical bone mineral apposition rate at left tibia of the periosteal surface, scale bar = 60 um F. Image of trabecular bone mineral apposition rate at L3 vertebra, scale bar = 40 um G. Comparison of cortical bone mineral apposition rates in left tibia among different groups (n = 7–8 per group) H. Comparison of trabecular bone mineral apposition rates at L3 vertebrae among different groups (n = 7–8 per group). DKK1-mAb: DKK1 monoclonal antibody, ALN: alendronate, SHAM: sham, HE: hematoxylin and eosin, TRAP staining: Tartrate-Resistant Acid Phosphatase staining: Identifies osteoclast activity in bone tissue, with TRAP-positive cells (osteoclasts) appearing red, indicating regions of active bone resorption, Von Kossa staining: Highlights areas of mineralization in bone tissue by staining calcium deposits black, Ct.MAR: cortical bone mineral apposition rate, Tb.MAR: trabecular bone mineral apposition rate ∗: P < 0.05, ∗∗: P < 0.01, ∗∗∗: P < 0.001 between the two groups.

Article Snippet: The embedded samples were sectioned into 10-μm-thick slices, de-plasticized, and stained with a Von Kossa stain kit (Servicebio, Cat# G1043).

Techniques: Staining, Comparison, Activity Assay

Journal: Materials Today Bio

Article Title: Deliver CEBPE via cartilage targeting Lipid nanoparticle to block CEBPE-LTF-STAT3 positive feedback loop for efficient treatment of cartilage endplate degeneration

doi: 10.1016/j.mtbio.2025.102027

Figure Lengend Snippet: CEBPE deficiency induces Inflammatory Responses, ECM degradation and calcification in EPCs. (A) WB analysis of CEBPE expression in EPCs that had been treated with the indicated doses and time of IL-1β (n = 3). (B) IF staining and quantification of CEBPE expression in EPCs treated with, or without, IL-1β (10 ng/mL) (n = 3) (Scale bar = 50 μm). (C) RT-qPCR analysis of the relative levels of CEBPE mRNA transcripts in EPCs treated with Si-NC, or with Si-CEBPE (n = 3). (D) WB analysis of CEBPE, IL-1β TNF expression in EPCs treated with Si-NC, or with Si-CEBPE (n = 3). (E) ELISA quantification of the levels of IL-1β and TNF in the supernatants of EPCs treated with Si-NC, or with Si-CEBPE (n = 3). (F) RT-qPCR analysis of the relative levels of Collagen II , MMP13 , RUNX2 , OPN , IL-1β , and TNF mRNA transcripts in EPCs treated with Si-NC, or with Si-CEBPE (n = 3). (G) WB analysis of Collagen II, MMP13, RUNX2, OPN expression in EPCs treated with Si-NC, or with Si-CEBPE (n = 3). (H, I) Representative IF staining of Collagen II, MMP13, and RUNX2 in EPCs treated with Si-NC, or with Si-CEBPE (Scale bar = 50 μm). (J) Representative Von Kossa staining in EPCs treated with Si-NC, or with Si-CEBPE (Scale bar = 100 μm). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: Cell calcification was assessed utilizing the Von Kossa Staining Kit from Solarbio (Cat. No: G3282).

Techniques: Expressing, Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Deliver CEBPE via cartilage targeting Lipid nanoparticle to block CEBPE-LTF-STAT3 positive feedback loop for efficient treatment of cartilage endplate degeneration

doi: 10.1016/j.mtbio.2025.102027

Figure Lengend Snippet: CEBPE overexpression alleviated IL-1β-induced the ECM degradation and calcification in EPCs. (A) RT-qPCR analysis of the relative levels of Collagen II , MMP13 , RUNX2 and OPN mRNA transcripts in OE-CEBPE or Empty Vector (EV) EPCs that had been pretreated with or without IL-1β (n = 3). (B) WB analysis of Collagen II, MMP13, RUNX2, and OPN expression in OE-CEBPE or EV EPCs that had been pretreated with or without IL-1β (n = 3). (C, D) Representative IF staining of Collagen II, MMP13, and RUNX2 in OE-CEBPE or EV EPCs that had been pretreated with or without IL-1β (Scale bar = 50 μm). (E) Representative Von Kossa staining in OE-CEBPE or EV EPCs that had been pretreated with or without IL-1β (Scale bar = 100 μm). n.s. not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: Cell calcification was assessed utilizing the Von Kossa Staining Kit from Solarbio (Cat. No: G3282).

Techniques: Over Expression, Quantitative RT-PCR, Plasmid Preparation, Expressing, Staining

Journal: Materials Today Bio

Article Title: Deliver CEBPE via cartilage targeting Lipid nanoparticle to block CEBPE-LTF-STAT3 positive feedback loop for efficient treatment of cartilage endplate degeneration

doi: 10.1016/j.mtbio.2025.102027

Figure Lengend Snippet: The feedback loop between CEBPE and the LTF/JAK2/STAT3 pathway in the ECM degradation and calcification of EPCs. (A) Experimental grouping diagram. (B) ELISA quantification of the levels of IL-1β and TNF in the supernatants of Si-NC or Si-CEBPE EPCs that had been transduced with LTF -overexpressing plasmids (OE-LTF), or with EV (n = 3). (C) RT-qPCR analysis of the relative levels of IL-1β , TNF , Collagen II , MMP13 , RUNX2 , and OPN mRNA transcripts in Si-NC or Si-CEBPE EPCs that had been transduced with OE-LTF, or with EV (n = 3). (D, E) WB analysis of IL-1β, TNF, Collagen II, MMP13, RUNX2, and OPN expression in Si-NC or Si-CEBPE EPCs that had been transduced with OE-LTF, or with EV (n = 3). (F, G) Representative IF staining of Collagen II, MMP13, and RUNX2 in Si-NC or Si-CEBPE EPCs that had been transduced with OE-LTF, or with EV. (H) Representative Von Kossa staining in Si-NC or Si-CEBPE EPCs that had been transduced with Si-LTF, or with EV. (I, J) WB analysis of Collagen II, MMP13, RUNX2 and OPN expression in Si-NC or Si-CEBPE EPCs that had been treated with or without AG490 (n = 3). (K) Schematic diagram of three potential STAT3 binding sites (Site 1, Site 2 and Site 3) in the CEBPE promoter. (L) The binding relationship between STAT3 and CEBPE -promoter as examined by dual-luciferase reporter gene assay. (M) ChIP-PCR for p-STAT3 enrichment in the CEBPE promoter. n.s. not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: Cell calcification was assessed utilizing the Von Kossa Staining Kit from Solarbio (Cat. No: G3282).

Techniques: Enzyme-linked Immunosorbent Assay, Transduction, Quantitative RT-PCR, Expressing, Staining, Binding Assay, Luciferase, Reporter Gene Assay