recombinant postn protein  (MedChemExpress)

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

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: Fibroblast-derived POSTN is highly expressed in obesity-related CRC. A . Differential expression analysis of POSTN in colorectal adenocarcinoma (COAD) samples versus normal samples from TCGA. B . Differential expression analysis of POSTN between paired adjacent normal and COAD tumor tissues from TCGA-COAD patients. C . Expression analysis of POSTN across different disease stages (I–IV) in TCGA-COAD patients. D . Survival analysis of TCGA-COAD patients by POSTN expression (low, n = 225; high, n = 225). The log-rank test was used for significance. E . Correlation analysis between POSTN expression and stromal scores in TCGA-COAD, assessed by Pearson correlation. F . Differential expression analysis of POSTN in WT-LF versus WT-HF groups from the GSE46843 dataset. G . Differential expression analysis of POSTN in APC (min/+) RD versus APC (min/+) HFD groups from the GSE278785 dataset. H . Representative IHC images of POSTN in normal colorectal tissues and COAD tumor tissues from obesity-associated CRC patients, with staining intensity categorized as weak or strong. Scale bars, 50 µm. The right panel quantifies the percentage of positive POSTN staining in normal, weak, and strong staining groups.I-L. scRNA-seq analysis of GSE231559 . ( I ) UMAP plot showing cell subtypes in the dataset: tumor cells, T cells, B cells, myeloid cells, fibroblasts, and endothelial cells. ( J ) Violin plot of POSTN expression across different cell subtypes (tumor cells, T cells, B cells, myeloid cells, fibroblasts, endothelial cells) in normal and tumor tissues. ( K ) UMAP plot showing fibroblast subtypes: fibroblasts, myCAFs, and iCAFs. ( L ) Violin plot of POSTN expression in fibroblast subtypes (fibroblasts, myCAFs, iCAFs) in normal and tumor tissues. M . Representative immunofluorescence co-staining of POSTN (green) and α-SMA (red) in normal and tumor tissues; Nuclei were counterstained with DAPI (blue). Scale bars, 50 µm. The right panels show intensity profiles of α-SMA and POSTN across lines drawn through the images for normal and tumor tissues.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Derivative Assay, Quantitative Proteomics, Expressing, Staining, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: POSTN promotes CRC occurrence and progression under high-fat diet. A . HFD-AOM-DSS induced CRC tumorigenesis protocol in WT (n = 10) and POSTN-KO (n=10) mice. Mice were fed HFD until sacrifice, injected with AOM (12.5mg/kg), followed by three cycles of 2% DSS (5 days per cycle, with 14-day intervals between cycles), and sacrificed at 72 days after the first DSS treatment. B . Representative images of colons from WT and POSTN-KO mice following HFD-AOM-DSS treatment. Scale bars, 1 cm. C . Quantification of body weight, colorectal length, tumor number, and tumor size in WT and POSTN-KO mice following HFD-AOM-DSS induction. D . Representative HE and Ki67 IHC images of colon tissues from WT and POSTN-KO mice after HFD-AOM-DSS treatment. Scale bars, 50 µm. E .Schematic diagram of the mice transplanted tumor model. Mice were fed RD or HFD for 12 weeks, then injected subcutaneously with mouse colorectal cancer cell line MC38, and sacrificed 21 days after injection. F . The volumes of subcutaneous transplanted tumors from the WT group and POSTN-KO group fed RD or HFD. G . Representative images of subcutaneous transplanted tumors from the WT group (n = 5) and POSTN-KO group (n= 5) fed RD or HFD. H .The size and weights of subcutaneous transplanted tumors in the WT group and POSTN-KO group fed RD or HFD. I . Representative HE and Ki67 IHC images of transplanted tissues from WT and POSTN-KO mice fed RD or HFD. Scale bars, 50 µm. J . Representative IHC images of POSTN, α-SMA and Collagen Ⅰ in colon tissues from WT and POSTN-KO mice induced by HFD-AOM-DSS. Scale bars, 50 µm. K . Representative IHC images of α-SMA and Collagen I in t subcutaneous transplanted tumors from WT and POSTN-KO mice fed RD or HFD. Scale bars, 50 µm. The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Injection

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: POSTN promotes fibroblast activation and promotes CRC development under high-fat diet. A . Representative IF images of biomarkers in primary WT and POSTN-KO fibroblasts. The merged images represent the overlay of DAPI (blue) with POSTN (red), α-SMA (red) and Collagen Ⅰ (red) respectively. Scale bars, 50 µm. B . Western blot analysis of POSTN, α-SMA, and Collagen Ⅰ expression levels in primary fibroblasts. Quantification of relative protein levels is shown adjacent to the blot images. C . Schematic diagram illustrating the mouse transplanted tumor model. Primary fibroblasts were isolated. Mice were fed a HFD for 12 weeks, then subcutaneously co-inoculated with MC38 cells along with WT or POSTN-KO primary fibroblasts to establish transplanted tumors. Mice were euthanized on day 21 post-inoculation for tumor harvest and analysis. D . Growth curves of subcutaneous transplanted tumors volumes in the MC38 + WT NFs group, and MC38 + POSTN-KO NFs group over 21 days. E . Representative images of subcutaneous transplanted tumors from the MC38+WT NFs group (n = 5) and MC38+KO NFs group (n = 5). Scale bar, 1 cm. F . Quantification of subcutaneous transplanted tumors size (left) and weight (right) in the MC38 + WT NFs group, and MC38 + POSTN-KO NFs group. G . Representative IHC images of α-SMA and Collagen I in subcutaneous transplanted tumors from the MC38+WT NFs group and MC38+POSTN-KO NFs group. Scale bars, 50 µm. Quantification of the percentage of positive staining for α-SMA and Collagen Ⅰ is shown adjacent to the IHC images. H .Western blot analysis and quantification of POSTN, STAT3 and p-STAT3 expression levels in WT primary fibroblasts treated with FFA. I .Western blot analysis and quantification of POSTN, STAT3, and p-STAT3 expression levels in WT primary fibroblasts treated with NSC74859, with or without FFA co-treatment.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Activation Assay, Western Blot, Expressing, Isolation, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: POSTN promotes lipid accumulation by regulating SREBP1 and CPT1A reprogramming under high-fat diet. A .GSEA results of mitochondrial fatty acid beta oxidation signaling pathway in TCGA-COAD patients. B . In the HFD-AOM-DSS model, morphology of mitochondria under transmission electron microscopy in colon tissues of WT and POSTN-KO mice. C . In HFD-AOM-DSS model, TG and NEFA levels in serum and colorectal tissues of WT and POSTN-KO mice. D . Intracellular TG and NEFA levels in MC38 cells following co-culture with WT or POSTN-KO NFs. E . Intracellular TG and NEFA levels in MC38 cells treated with DMEM, WT-CM (conditioned medium from WT NFs), or KO-CM (conditioned medium from POSTN-KO NFs). F . Neutral lipid accumulation in MC38 cells treated with DMEM, WT-CM, or KO-CM, visualized by BODIPY staining (green, neutral lipid; blue, nuclei). Scale bars, 50 μm. G . Intracellular TG and NEFA levels in MC38 cells treated with WT-CM, KO-CM, or WT-CM plus orlistat. H . Cell viability of MC38 cells treated with WT-CM, KO-CM, or WT-CM plus orlistat. I . Neutral lipid accumulation in MC38 cells treated with WT-CM, KO-CM, or WT-CM plus orlistat, visualized by BODIPY staining. Scale bars, 50 μm. J . Western blot analysis of SREBP1 and CPT1A expression levels in MC38 cells treated with WT-CM or KO-CM when FFA was added. Quantification of relative protein levels is shown adjacent to the blot images. K . Western blot analysis of SREBP1 expression levels in MC38 cells transfected with empty vectors or SREBP1 overexpression plasmids under KO-CM treatment. Quantification of relative protein levels is shown adjacent to the blot images.Right figure: BODIPY staining of neutral lipid in cells. Scale, 50 μm. L . Western blot analysis of CPT1A expression levels in MC38 cells transfected with empty vectors or CPT1A overexpression plasmids under WT-CM treatment. Quantification of relative protein levels is shown adjacent to the blot images. Right figure: BODIPY staining of neutral lipid in cells. Scale, 50 μm.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Transmission Assay, Electron Microscopy, Co-Culture Assay, Staining, Western Blot, Expressing, Transfection, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: POSTN promotes FA synthesis via the AKT/SREBP1 signaling axis under high-fat diet. A . GSEA of the REACTOME_PI3K_AKT_SIGNALING_IN_CANCER pathway in TCGA-COAD patients. Genes are ranked by correlation with POSTN expression (red, POSTN-high; green, POSTN-low). B . Western blot analysis of AKT, p-AKT, and SREBP1 expression levels in tissues from WT and POSTN–KO mice subjected to HFD-AOM-DSS induction. Quantification of relative protein levels is shown adjacent the blot images. C .Western blot analysis of AKT and p-AKT expression levels in HCT116 and HT29 cells treated with FFA with or without recombinant POSTN supplementation. Quantification of relative protein levels is shown below the blot images. D . Western blot analysis of AKT, p-AKT, and SREBP1 expression levels in HCT116 and HT29 cells treated with FFA, with or without the AKT inhibitor LY294002 and/or recombinant POSTN supplementation. Quantification of relative protein levels is shown below to the blot image. E .Intracellular TG and NEFA levels in HCT116 and HT29 cells treated with FFA, with or without LY294002 and/or POSTN supplementation. F . Representative fluorescence microscopy images of neutral lipid accumulation (green, BODIPY staining) in HCT116 and HT29 cells treated with FFA, with or without LY294002 and/or POSTN supplementation. Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Expressing, Western Blot, Recombinant, Fluorescence, Microscopy, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: POSTN inhibits FAO via the ERK/CPT1A signal axis under high-fat diet. A . GSEA of the REACTOME_ONCOGENIC_MAPK_SIGNALING pathway in TCGA-COAD patients. Genes are ranked by correlation with POSTN expression (red, POSTN-high; green, POSTN-low). B .Western blot analysis of ERK, p-ERK, and CPT1A expression levels in tissues from WT and POSTN-KO mice subjected to HFD-AOM-DSS to induce. Quantification of relative protein levels is shown adjacent the blot images. C .Western blot analysis of ERK and p-ERK expression levels in HCT116 and HT29 cells treated with FFA with or without recombinant POSTN supplementation. Quantification of relative protein levels is shown below the blot images. D .Western blot analysis of ERK, p-ERK, and CPT1A expression levels in HCT116 and HT29 cells treated with FFA, with or without the ERK inhibitor PD98059 and/or recombinant POSTN supplementation. Quantification of relative protein levels is shown below to the blot image. E .Intracellular TG and NEFA levels in HCT116 and HT29 cells treated with FFA, with or without PD98059 and/or POSTN supplementation. F .Representative fluorescence microscopy images of neutral lipid accumulation (green, BODIPY staining) in HCT116 and HT29 cells treated with FFA, with or without PD98059 and/or POSTN supplementation. Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Expressing, Western Blot, Recombinant, Fluorescence, Microscopy, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: Cilengitide targeted therapy for CRC associated with high-fat diet. A .Western blot analysis of AKT, p-AKT, SREBP1, ERK, p-ERK, and CPT1A in HCT116 and HT29 cells treated with recombinant POSTN with or without Cilengitide. Quantification of relative protein levels is shown adjacent to the blot images. B . Intracellular TG and NEFA levels in HCT116 and HT29 cells treated with recombinant POSTN with or without Cilengitide. C . Representative fluorescence microscopy images of neutral lipid accumulation (green, BODIPY staining) in HCT116 and HT29 cells treated with recombinant POSTN with or without Cilengitide. Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm. D .The volumes of subcutaneous transplanted tumors from the control group, LY294002 group, PD98059 group, Cilengitide group and Cilengitide+LY294002+PD98059 group. E . Representative images of subcutaneous transplanted tumors from control group (n = 5), LY294002 group (n = 5), PD98059 group (n = 5), Cilengitide group (n = 5), and Cilengitide+LY294002+PD98059 group (n = 5). F . The size and weights of subcutaneous transplanted tumors from the control group, LY294002 group, PD98059 group, Cilengitide group, and Cilengitide+LY294002+PD98059 group. G . Representative HE and IHC images of Ki-67 and α-SMA of subcutaneous transplanted tumors from the control group, LY294002 group, PD98059 group, Cilengitide group, and Cilengitide+LY294002+PD98059 group. Scale bars, 50 µm. H .Western blot analysis of AKT, p-AKT, SREBP1, ERK, p-ERK, and CPT1A of subcutaneous transplanted tumors from the control group, LY294002 group, PD98059 group, Cilengitide group, and Cilengitide+LY294002+PD98059 group. Quantification of relative protein levels is shown adjacent to the blot images.The data are means ± SD, for all panels: * P < 0.05; ** P < 0.01

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques: Western Blot, Recombinant, Fluorescence, Microscopy, Staining, Control

Journal: Cell Communication and Signaling : CCS

Article Title: Fibroblast-derived POSTN promotes colorectal cancer progression under high-fat diet by reprogramming fatty acid metabolism in tumor cell

doi: 10.1186/s12964-025-02618-w

Figure Lengend Snippet: Schematic diagram of POSTN regulating FA metabolism in colorectal cancer cells with a high-fat diet

Article Snippet: The recombinant protein POSTN (100 ng/mL, 10299-H08H, Sinobiological), AKT inhibitor LY294002 (20 μM, L832989-10 mg, Macklin), ERK inhibitor PD98059 (10μM, P832941-5 mg, Macklin), Cilengitide (10 μM, BD628662, Bidepharm) was applied to HCT116 and HT29 cells for 24 h. MC38 cells were treated with CM.

Techniques:

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: Periostin mediates the pro-fibrogenic activity of Adam12 + fibroblasts during skin scarring. A Schematic depicting wounding and tamoxifen induction of Adam12 CreERT2 ;Postn fl/fl mice for Adam12 + cells-specific Postn deletion. B, C Immunofluorescence staining analysis for periostin on wound tissues of Adam12 CreERT2 ;Postn +/+ , Adam12 CreERT2 ;Postn fl/+ , and Adam12 CreERT2 ;Postn fl/fl mice at 14dpw. Error bars represent SD (n=8-16 wounds). Scale bar = 100μm. D, E Masson staining on wound tissues of Adam12 CreERT2 ;Postn +/+ , Adam12 CreERT2 ;Postn fl/+ , and Adam12 CreERT2 ;Postn fl/fl mice at 14dpw. Error bars represent SD (n=8-16 wounds). Scale bar = 100μm. F Schematic depicting wounding, 2mg tamoxifen induction, 200ng DT injection and 1μg rmPOSTN (rmPN) injection of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. G, H Immunofluorescence staining analysis for periostin on wound tissues of PBS group and DT group Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice at 14dpw. Error bars represent SD (n=5 wounds). Scale bar = 100μm. I, J Representative photographic images of wound tissues of PBS group, DT group and DT+rmPOSTN group Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice at 0dpw and 14dpw. Error bars represent SD (n=5 wounds). K, L Masson staining on wound tissues of PBS group, DT group and DT+rmPOSTN group Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice at 14dpw. Error bars represent SD (n=5 wounds). Scale bar = 100μm. One-way ANOVA test was used to determine statistical significance in C, E, J and L . Two-tailed Student's unpaired t-test was used to determine statistical significance in H .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Activity Assay, Immunofluorescence, Staining, Injection, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: BMP2 signaling is required for the lineage commitment of resident dermal fibroblasts into pro-fibrogenic Adam12 + fibroblasts. A Schematic depicting wounding and tamoxifen induction of Pdgfra CreERT2 ;Bmpr2 fl/fl mice for fibroblasts-specific Bmpr2 deletion. B, C Immunofluorescence staining analysis for Adam12 on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. D, E Representative photographic images of wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 0dpw and 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). F, G Masson staining on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. H, I Immunofluorescence staining analysis for periostin on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. One-way ANOVA test was used to determine statistical significance in C, E, G and I .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Immunofluorescence, Staining

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: Augmentation of BMP2 signaling drives expansion of Adam12 + fibroblasts and exacerbates skin scarring. A-F, Immunofluorescence staining analysis for BMP2, ADAM12 and Periostin in normal scar, hypertrophic scar (HTS) and keloid. Error bars represent SD (n=8). Scale bar = 100μm. G The correlation analyses between the expression of BMP and ADAM12. The range between the two dashed lines represents the 95% confidence interval. H The correlation analyses between the expression of ADAM12 and periostin. The range between the two dashed lines represents the 95% confidence interval. I Schematic depicting wounding, 2mg tamoxifen induction, 200ng DT injection and 25ng BMP2 injection of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. J, K Flow cytometry analysis of tdTomato on vehicle+PBS injection, BMP2+PBS injection and BMP2+DT injection wounds of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. Error bars represent SD (n=6 wounds). L, M Representative photographic images of wound tissues of vehicle+PBS group, BMP2+PBS group and BMP2+DT group Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice at 0dpw and 14dpw. Error bars represent SD (n=6 wounds). N, O Masson staining on vehicle+PBS injection, BMP2+PBS injection and BMP2+DT injection wounds of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. Error bars represent SD (n=6 wounds). Scale bar = 100μm. N Schematic illustration showing that abnormal high-expressed BMP2 in healed wound may lead to pathologic scars. One-way ANOVA test was used to determine statistical significance in B, D, F, K, M and O . Pearson correlation was used to measures the strength and direction of a linear relationship between two variables in G and H .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Immunofluorescence, Staining, Expressing, Injection, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: BMP2 represents a promising therapeutic target for pathological scars. A Schematic depicting wounding, vehicle injection and 20μg BMP2 inhibitor LDN-193189 2HCl injection of tension induced hypertrophic scar mice model. B Representative photographic images of scar tissues of vehicle injection or LDN-193189 2HCl injection group in tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 5mm. C, D Immunofluorescence staining analysis for Adam12 on vehicle injection or LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. E, F Masson staining on vehicle injection and LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. G-L, Immunofluorescence staining analysis for Collagen I, Collagen III and periostin on vehicle injection and LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. Two-tailed Student's unpaired t-test was used to determine statistical significance in D, F, H, J and L .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Injection, Immunofluorescence, Staining, Two Tailed Test

Journal: Cancer Research Communications

Article Title: Periostin Promotes Sarcoma Growth by Promoting Tumor-Associated Macrophage Migration and Differentiation

doi: 10.1158/2767-9764.CRC-25-0301

Figure Lengend Snippet: Postn silencing reduces tumor growth in the p53 KO Vgll3 OE mouse model. A, Western blot analysis of POSTN expression in murine Postn-WT and Postn-OE sarcoma cells (p53 KO Vgll3 OE ). B, Soft agar assays comparing Postn-WT and Postn-OE sarcoma cells (p53 KO Vgll3 OE ). Representative images of the analyzed spheroids (right) and spheroid area quantification (left) are shown. Each dot represents a quantified imaging area. C, Relative expression of Postn in FACS-sorted tumor cells upon Postn silencing through shRNAs (shPostn) in p53 KO Vgll3 OE sarcoma cells. D, Tumor size at different time points, comparing Postn-WT (shSCR) and Postn-silenced (shPostn) sarcoma cells (p53 KO Vgll3 OE ). E, Weight of the tumor masses at the experimental endpoint, comparing tumors generated by Postn-WT (shSCR) and Postn-silenced (shPostn) sarcoma cells (p53 KO Vgll3 OE ). F, Heatmap of the top differentially expressed genes with absolute log 2 fold change > 1. G, Top pathways enriched in shPostn vs. shSCR sarcoma cells (p53 KO Vgll3 OE ). No pathways were enriched in shSCR vs. shPostn tumor cells. Unless otherwise indicated, results are presented as mean ± SEM, and P values were derived from a two-tailed unpaired Student t test (*, P < 0.05; **, P < 0.01). All experiments were performed in the p53 KO Vgll3 OE sarcoma model. WT, wild type.

Article Snippet: Cells at different time points were collected for Transwell assays or treatment with recombinant POSTN (2955-F2-050, R&D Systems).

Techniques: Western Blot, Expressing, Imaging, Generated, Derivative Assay, Two Tailed Test

Journal: Cancer Research Communications

Article Title: Periostin Promotes Sarcoma Growth by Promoting Tumor-Associated Macrophage Migration and Differentiation

doi: 10.1158/2767-9764.CRC-25-0301

Figure Lengend Snippet: POSTN expression is correlated with patient survival and expression of extracellular matrix proteins. A, Kaplan–Meier plot showing reduced survival for patients with sarcoma expressing high POSTN levels. B and C, Pathway analysis of patients with sarcoma showing high vs. low POSTN expression levels revealed increased expression of ECM-related elements in POSTN-high patients. D, Schematic representation of the sarcoma mouse models. E, Volcano plot of select differentially expressed genes in tumor cells sorted from the p53 KO Vgll3 OE (positive log fold change) vs. p53 KO Ccne1 OE (negative log fold change) mouse tumors. F, Heat map showing the most differentially expressed soluble factors in the ELISA array comparing p53 KO Vgll3 OE vs. p53 KO Ccne1 OE whole tumors. G, Expression of Postn mRNA in tumor cells, adjacent CAFs, and bone marrow–derived mesenchymal cells (CD45 − PDGFRA + ). Unless otherwise indicated, results are presented as mean ± SEM, and P values were derived from a two-tailed unpaired Student t test (*, P < 0.05). [Created in BioRender. Guarnerio, J. (2025) https://BioRender.com/fo00cpj .]

Article Snippet: Cells at different time points were collected for Transwell assays or treatment with recombinant POSTN (2955-F2-050, R&D Systems).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Two Tailed Test

Journal: Cancer Research Communications

Article Title: Periostin Promotes Sarcoma Growth by Promoting Tumor-Associated Macrophage Migration and Differentiation

doi: 10.1158/2767-9764.CRC-25-0301

Figure Lengend Snippet: Postn silencing in tumor cells shapes the sarcoma immune microenvironment. A, Flow cytometry analysis of CD45 + immune cells and different types of immune cells within the CD45 + population in the microenvironment of Postn-WT (shSCR) and Postn-silenced (shPostn) tumors (p53 KO Vgll3 OE ). Each dot represents one mouse. B, Subclustering of the mononuclear phagocyte compartment (macrophages, monocytes, and dendritic cells) from scRNA-seq. C, Proportions of the subclusters from B in individual shSCR or shPostn tumors ( n = 5 mice in each group). D, Pathways enriched in mononuclear phagocytes from shPostn tumors (top) or from shSCR tumors (bottom). E, Relative expression of IFN-stimulated and proinflammatory genes in CD11b + myeloid cells FACS-sorted from mouse tumors, generated by either Postn -WT (shSCR) or Postn -silenced (shPostn) sarcoma cells. Each dot represents one mouse. F, Flow cytometry analysis of CD45 + immune cells and different types of immune cells within the CD45 + population in the microenvironment of mice treated with POSTN-blocking antibody (anti-POSTN). Each dot represents one mouse. Unless otherwise indicated, results are presented as mean ± SEM, and P values were derived from a two-tailed unpaired Student t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001). Experiments were performed in the p53 KO Vgll3 OE model.

Article Snippet: Cells at different time points were collected for Transwell assays or treatment with recombinant POSTN (2955-F2-050, R&D Systems).

Techniques: Flow Cytometry, Expressing, Generated, Blocking Assay, Derivative Assay, Two Tailed Test

Journal: Cancer Research Communications

Article Title: Periostin Promotes Sarcoma Growth by Promoting Tumor-Associated Macrophage Migration and Differentiation

doi: 10.1158/2767-9764.CRC-25-0301

Figure Lengend Snippet: Postn recruits bone marrow monocytes and promotes the accumulation of macrophages. A, Quantification of POSTN levels in the serum of sarcoma-bearing mice compared with healthy mice. Each dot represents one mouse. B, Quantification of POSTN levels in the bone marrow of sarcoma-bearing mice compared with healthy mice. Each dot represents one mouse. C, Transwell migration assays to measure the capability of bone marrow–derived monocytes to migrate toward recombinant POSTN protein after 16 hours of exposure or ( D ) in the presence of POSTN-neutralizing antibodies. E, Transwell migration assays to assess the capability of bone marrow–derived monocytes to migrate toward supernatant from sarcoma cells expressing wild-type (Postn-WT) or enhanced Postn levels (Postn-OE). F and G, Analysis of monocytes (Ly6C+F4/80 − cells) and macrophages (Ly6C-F4/80 + cells) at two different time points (days 4 and 6) in culture with recombinant POSTN protein. Unless otherwise indicated, results are presented as mean ± SEM, and P values are derived from a two-tailed unpaired Student t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001). For C–G , each dot represents BMDM derived from a single mouse ( n = 4 independent mice). [Created in BioRender. Guarnerio, J. (2025) https://BioRender.com/fo00cpj .]

Article Snippet: Cells at different time points were collected for Transwell assays or treatment with recombinant POSTN (2955-F2-050, R&D Systems).

Techniques: Migration, Derivative Assay, Recombinant, Expressing, Two Tailed Test