Journal: Scientific Reports

Article Title: Integrated bioinformatic analysis and experiments reveal EFEMP1 as a novel aging-related signature gene in calcific aortic valve disease

doi: 10.1038/s41598-026-45986-0

Figure Lengend Snippet: ScRNA-seq analysis. A UMAP displaying the different cell types. B Dotplot displaying the markers used for identifying cell types. C-F Feature plot and violin plot displaying the distribution of Il6 and Efemp1 among different cell types. G-H The expression of Il6 and Efemp1 in VICs. I-J Feature plot and violin plot displaying the distribution of EFEMP1 among different cell types. K The EFEMP1 expression in VICs.

Article Snippet: Following deparaffinization and antigen retrieval, sections underwent immunohistochemical staining using a primary anti-EFEMP1 antibody (HUABIO, Cat# ER63434, 1:300 dilution).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Integrated bioinformatic analysis and experiments reveal EFEMP1 as a novel aging-related signature gene in calcific aortic valve disease

doi: 10.1038/s41598-026-45986-0

Figure Lengend Snippet: Expression signature and ROC analysis. A-C Box plot showing the expression signature of EFEMP1. D-F ROC curves displaying the diagnostic value of EFEMP1.

Article Snippet: Following deparaffinization and antigen retrieval, sections underwent immunohistochemical staining using a primary anti-EFEMP1 antibody (HUABIO, Cat# ER63434, 1:300 dilution).

Techniques: Expressing, Diagnostic Assay

Journal: Scientific Reports

Article Title: Integrated bioinformatic analysis and experiments reveal EFEMP1 as a novel aging-related signature gene in calcific aortic valve disease

doi: 10.1038/s41598-026-45986-0

Figure Lengend Snippet: Experimental validation of EFEMP1 expression in human aortic valve. A Macroscopic (scale bar: 100 μm) and micrographs (scale bar: 25 μm) views of immunohistochemical staining in human aortic valve tissue sections, and histograms quantifying immunohistochemical staining results (n = 7). B Immunofluorescent staining of EFEMP1 and Vimentin in aortic valves (scale bar: 25 μm).

Article Snippet: Following deparaffinization and antigen retrieval, sections underwent immunohistochemical staining using a primary anti-EFEMP1 antibody (HUABIO, Cat# ER63434, 1:300 dilution).

Techniques: Biomarker Discovery, Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Integrated bioinformatic analysis and experiments reveal EFEMP1 as a novel aging-related signature gene in calcific aortic valve disease

doi: 10.1038/s41598-026-45986-0

Figure Lengend Snippet: Experimental validation of EFEMP1 expression at cellular level and functional experiment. A Alizarin Red staining after osteogenic induction for 14 days (scale bar: 100 μm). B The qRT-PCR results of EFEMP1, RUNX2, BMP2 and ALP in VICs induced by osteogenic medium(n = 6). C Representative Western blot analysis and quantification of EFEMP1 protein levels in VICs induced by osteogenic medium(n = 6) . D Representative Western blotting analysis and quantification of protein levels in the EFEMP1 knockdown experiment(n = 3).

Article Snippet: Following deparaffinization and antigen retrieval, sections underwent immunohistochemical staining using a primary anti-EFEMP1 antibody (HUABIO, Cat# ER63434, 1:300 dilution).

Techniques: Biomarker Discovery, Expressing, Functional Assay, Staining, Quantitative RT-PCR, Western Blot, Knockdown

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Effect of EFEMP1 knockdown by small interfering RNA (siRNA) on chondrocyte phenotype. A The human chondrocyte cell line C20A4 was transfected with either EFEMP1 siRNA (siEFEMP1) or scrambled siRNA (siCtrl). EFEMP1 and MMP-13 protein expression was analyzed by Western blotting. B Chondrocyte-related gene expression, including SOX9, aggrecan (ACAN), MMP13, type X collagen (COL10A1), and type II collagen (COL2A1), was assessed by quantitative PCR in siEFEMP1- and siCtrl-treated C20A4 cells. C Phosphokinase protein arrays were used to profile phosphorylation changes in intracellular proteins extracted from C20A4 chondrocytes. D Spot intensities exhibiting changes greater than ± 25% between groups were quantified using ImageJ software. Red lines indicate baseline levels from the siCtrl group. E Phosphorylation of p70S6K at threonine 421/serine 424 (T421/T424) was further examined by Western blot analysis. Data are presented as mean ± standard error of the mean (SEM). Statistical significance was determined using an independent-samples t-test. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Knockdown, Small Interfering RNA, Transfection, Expressing, Western Blot, Gene Expression, Real-time Polymerase Chain Reaction, Phospho-proteomics, Software

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Effect of EFEMP1 knockdown on osteoblast differentiation and mineralization in preosteoblastic cells. A The preosteoblastic MC3T3-E1 cell line was transfected with either EFEMP1 siRNA (siEFEMP1) or scrambled siRNA (siCtrl). EFEMP1 and RUNX2 protein expression was analyzed by Western blotting. B, C Alizarin Red staining was used to assess matrix mineralization in MC3T3-E1 cells cultured in osteogenic induction medium (OIM), with quantification performed on days 21, 28, and 35. D Alkaline phosphatase (ALP) staining was performed to evaluate osteogenic differentiation. E ALP enzymatic activity was measured in cell lysates on days 9 and 14 following siRNA transfection. F Osteopontin (OPN) expression was analyzed on days 3 and 7 during osteogenic induction. Data are presented as mean ± standard error of the mean (SEM). Statistical analyses: Data in A were analyzed using an independent-samples t -test, while data in C , E , and F were analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Knockdown, Transfection, Expressing, Western Blot, Staining, Cell Culture, Activity Assay, Comparison

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: EFEMP1 antibody treatment and cartilage histological assessment in a mouse model of osteoarthritis (OA). A Schematic illustration of the EFEMP1 antibody treatment protocol in STR/ort mice from 16 to 21 weeks of age. B Representative Safranin O–stained knee joint sections, with magnified views of the non-calcified cartilage (NCC) and calcified cartilage (CC) regions. Matrix-producing chondrocytes (MPCs) and matrix-non-producing chondrocytes (MNCs) are indicated by arrows. C Quantification of MPC and MNC numbers in the NCC and CC regions of articular cartilage, with counts combined from medial and lateral compartments. D Osteoarthritis severity was evaluated using Osteoarthritis Research Society International (OARSI) scores across four joint compartments: lateral femoral condyle (LF), medial femoral condyle (MF), lateral tibial plateau (LT), and medial tibial plateau (MT). Data in C are presented as mean ± standard error of the mean (SEM), while data in D are presented as median and quartiles using a violin plot. Statistical analysis was performed using an independent-samples t-test. * P < 0.05 and ** P < 0.01

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Staining

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Immunofluorescence analysis of cartilage markers following EFEMP1 antibody treatment in STR/ort mice. A Representative immunofluorescence (IF) staining for EFEMP1, SOX9, MMP13, aggrecan fragments, COL2A1, and COL10A1 in the medial femoral condyle (MF) and medial tibial plateau (MT). Nuclei were counterstained with DAPI (blue), and articular cartilage regions are outlined by orange dashed lines. B Quantification of fluorescence intensity in articular cartilage regions of the femoral condyle and tibial plateau, with medial and lateral compartments combined. EFEMP1 and SOX9 signals were normalized to DAPI, whereas MMP13, aggrecan fragments, COL10A1, and COL2A1 signals were quantified as absolute FITC intensity. Data are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using an independent-samples t -test. P < 0.05, P < 0.01, and ** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Cytokine profiling following EFEMP1 antibody treatment in STR/ort mice with osteoarthritis. A Heat-map representation of serum cytokine expression profiles measured using a multiplex assay. B Quantification of serum cytokine concentrations determined by Luminex xMAP analysis. Data are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test. P < 0.05, P < 0.01, and P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Expressing, Multiplex Assay, Luminex