Journal: BMC Veterinary Research

Article Title: Comparative analysis of the surface exposed proteome of two canine osteosarcoma cell lines and normal canine osteoblasts

doi: 10.1186/1746-6148-9-116

Figure Lengend Snippet: Quantitative real-time PCR of cell lines. Quantitative real-time PCR indicating relative expression of selected genes corresponding to surface proteins detected by mass spectrometry of cultured normal canine osteoblasts (CnOb) and two validated canine osteosarcoma cell lines (POS and HMPOS).

Article Snippet: The canine osteosarcoma cell lines POS and HMPOS [ ] were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and the normal canine osteoblast cell line, CnOb (Cell Applications, San Diego, CA), was cultured in canine osteoblast medium (Cell Applications, San Diego, CA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Mass Spectrometry, Cell Culture

Journal: BMC Veterinary Research

Article Title: Comparative analysis of the surface exposed proteome of two canine osteosarcoma cell lines and normal canine osteoblasts

doi: 10.1186/1746-6148-9-116

Figure Lengend Snippet: Western blot of cell lines. Western blot of whole cell lysate to detect the presence of CD44, Thrombospondin-1 and CYR61 in cultured normal canine osteoblasts (CnOb) and two validated canine osteosarcoma cell lines (POS and HMPOS).

Article Snippet: The canine osteosarcoma cell lines POS and HMPOS [ ] were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and the normal canine osteoblast cell line, CnOb (Cell Applications, San Diego, CA), was cultured in canine osteoblast medium (Cell Applications, San Diego, CA).

Techniques: Western Blot, Cell Culture

Journal: PLoS ONE

Article Title: Demineralized bone matrix used for direct pulp capping in rats

doi: 10.1371/journal.pone.0172693

Figure Lengend Snippet: (A): Representative immunohistochemical images of the blank control group, the Ca(OH) 2 group and the DBM group. a: The blank control group. Pulp morphology was normal. b- f: Ca(OH) 2 group (1, 3, 7, 14, 28 days). g- k: DBM group (1, 3, 7, 14, 28 days). (B): Mean IOD value of Runx2. *means significant difference.

Article Snippet: The sections were deparaffinized with xylene, hydrated in a series of descending grades of ethanol, and then rinsed briefly with PBS for the primary antibodies; the sections were incubated overnight at 4°C with polyclonal anti- Runx2, COL I, OCN and DSP (Wuhan Boster Biological Technology, Wuhan, China).

Techniques: Immunohistochemical staining, Control

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN in M1 macrophage‐derived SFs mediates the formation of traumatic HO. A) High‐throughput sequencing was performed between SFs derived from macrophages and M1‐macrophages in vitro and between the sham group and the tendon lesions at 7 days in vivo. An intersection Venn diagram was drawn. B) WB analysis was used to detect the expression of POSTN proteins in Mφ‐SFs, M1‐SFs, and NFκB knock out groups, N = 3, ** p < 0.01, *** p < 0.001, **** p < 0.0001. C) IHC staining was used to detect the expression of POSTN in the sham, positive, and NFκB knock‐out groups, N = 6. D) Immunofluorescence staining for the Runx2, OCN, OPN for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3. E) WB analysis was used to detect the expression of osteogenic‐related protein levels (Runx2, OCN, OPN) for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3, **** p < 0.0001. F) ALP and ARS staining for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001. G) Immunofluorescence staining for the Runx2 of tendons and Micro‐CT analysis of HO formation in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Derivative Assay, Next-Generation Sequencing, In Vitro, In Vivo, Expressing, Knock-Out, Immunohistochemistry, Immunofluorescence, Staining, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN promotes the formation of traumatic HO by enhancing β‐oxidation of fatty acids. A) High‐throughput whole‐transcriptome sequencing was performed and showed by Reactome pathways enrichment analysis of RNA‐seq data between the sham group and tendon lesions at 7 days, N = 3 B) Immunofluorescence staining for LCAD and MCAD of TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups. C) WB analysis was used to detect the expression of LCAD and MCAD proteins for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3, **** p < 0.0001. D) Seahorse test was used to detect the oxidative phosphorylation level in the osteogenic induced TDSCs in addition of M1‐SFs or M1‐SFs with POSTN knockout groups, N = 3, ● represented M1‐SFs with POSTN knockout groups and ▲ represented M1‐SFs groups. E) Fluorescence and light microscope and WB analysis were used to confirm the success of downregulation of LCAD transfection for TDSCs, N = 3. F) Immunofluorescence staining for the LCAD and MCAD (red), co‐localized with PDGFRα(green) of tendons in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 3. G) WB analysis was used to detect the expression of LCAD and MCAD of TDSCs in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 3, **** p < 0.0001. H) ALP and ARS staining were used to detect the osteogenesis of TDSCs in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 6, **** p < 0.0001. I) Immunofluorescence staining for the Runx2 of tendons and micro‐CT analysis of HO formation in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: High Throughput Screening Assay, Sequencing, RNA Sequencing, Immunofluorescence, Staining, Knock-Out, Expressing, Phospho-proteomics, Fluorescence, Light Microscopy, Transfection, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN enhances osteogenic propensity by binding to PTK7. A) Mass spectrometry analysis was used and presented through a Venn diagram to detect the potential molecules that showed increased binding to POSTN in the disease model between the sham group and the tendon injury group at 7 days, N = 3. B) Mass spectrum of the binding between PTK7 and POSTN. C) According to the Score Sequest HT, PTK7 ranks first among the molecules that bind to POSTN in the heterotopic ossification model. D) Docking images showing the predicted binding position of POSTN and PTK7 protein. E) IF staining showed co‐localization of POSTN and PTK7 in the cytoplasm of TDSCs, N = 3. F) IF staining showed co‐localization of POSTN and PTK7 in the heterotopic ossified tissue, N = 6. G) Co‐IP analysis was used to verified the bind relationship between POSTN and PTK7 in the osteogenic induced TDSCs, N = 3. H) Co‐IP analysis was used to verified the bind relationship between POSTN and PTK7 in the heterotopic ossified tissue, N = 3. I) Immunofluorescence staining indicated that overexpression of PTK7 could upregulate the protein expression levels of LCAD and MCAD in the absence of POSTN in vitro, N = 3. J) ALP and ARS staining was used to detect the osteogenesis of TDSCs in the POSTN‐/‐ and POSTN‐/‐&OV‐PTK7 groups, N = 6, **** p < 0.0001. K) Immunofluorescence staining for the Runx2 of tendons and micro‐CT analysis of HO formation in the POSTN‐/‐ and POSTN‐/‐&OV‐PTK7 groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Binding Assay, Mass Spectrometry, Staining, Co-Immunoprecipitation Assay, Immunofluorescence, Over Expression, Expressing, In Vitro, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN promotes the osteogenic transition of TDSCs by mediating the phosphorylation of AKT at the S124 site. A) High‐throughput sequencing was performed between TDSCs treated with SFs derived from M1 macrophages and SFs derived from M1 macrophages with the POSTN protein knocked out respectively and between overexpressed phosphorylation at the AKT S124 site and the mutated S124 site respectively. An intersection Venn diagram was drawn. B) WB analysis was used to detect the expression of CPT1 proteins of TDSCs in the M1‐SFs, POSTN‐/‐, sh‐PTK7, and mut‐S124 groups, N = 3, **** p < 0.0001. C) Immunofluorescence staining for LCAD and MCAD of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 3. D) ALP and ARS staining were used to detect the osteogenesis of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 6, **** p < 0.0001. E) WB analysis was used to detect the expression of LCAD and MCAD proteins of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 3, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Phospho-proteomics, Next-Generation Sequencing, Derivative Assay, Expressing, Immunofluorescence, Staining